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© 1997 Oxford University Press 3564-3569

The Elav-like proteins bind to AU-rich elements and to the poly(A) tail of mRNA

The Elav-like proteins bind to AU-rich elements and to the poly(A) tail of mRNA Wei-Jun Ma, Sangmi Chung and Henry Furneaux*

Program in Molecular Pharmacology and Therapeutics, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY 10021, USA

Received June 25, 1997; Accepted July 15, 1997

ABSTRACT

The Elav-like proteins are specific mRNA binding proteins which are required for cellular differentiation. They contain three characteristic RNP2/RNP1-type RNA binding motifs. Previously we have shown that the first and second RNA binding domains bind to AU-rich elements in the 3'-UTR of mRNA. In this paper we show that the Elav-like proteins exhibit poly(A) binding activity. This activity is distinct from poly(A) binding activities that have been previously described. The Elav-like proteins specifically bind to long chain poly(A) tails. We have shown that the third RNA binding domain encompasses this poly(A) binding activity. Using poly(A)-Sepharose beads in a `sandwich' assay we have shown that the Elav-like proteins can bind simultaneously to the AU-rich element and to the poly(A) tail.

INTRODUCTION

The Elav-like proteins are a group of closely related RNA binding proteins that were first described in Drosophila (1 ). Recently, Elav-like genes have been cloned from higher organisms (2 -8 ). There are four members of the human family, HuD, HuC, Hel-N1 and HuR (3 -5 ,7 ). HuR is expressed at the RNA level in all proliferating cells, whereas HuD, HuC and Hel-N1 are normally expressed on terminal differentiation of neurons (7 ,9 ,10 ). The human members of this family are of particular interest since they are tumor antigens associated with a wide variety of human tumors. (5 ,11 ,12 ). The Elav-like proteins contain three highly conserved RNA recognition motifs (RRM). Two of these RRMs are in tandem and are separated from the third by a basic segment (3 ,4 ,7 ,13 ). A significant insight into the mechanism of action of these proteins was provided by the observation that they specifically bind to AU-rich elements in the 3'-untranslated region (UTR) of mRNA. (7 ,13 -17 ). These AU-rich elements were originally characterized by Shaw and Kamen, who were the first to show that the AU-rich element in the 3'-UTR of GM-CSF mRNA regulates its expression at the post-transcriptional level (18 ). The current model is that the AU-rich elements are recognized by a specific endonuclease, which cleaves the transcript and renders it acessible to an exonuclease (19 ). Thus, mRNAs that contain these elements have a very short half-life and are usually present at a very low steady-state level. The level of these mRNAs can be dramatically increased by factors that also bind to the AU-rich elements and inhibit the degradative activity. Recent evidence has indicated that the Elav-like proteins are such factors and selectively inhibit the decay of mRNAs that contain AU-rich elements (20 -23 ). In previous studies we have shown that the first and second RRMs of HuD and HuR bind specifically and with high affinity to AU-rich elements (7 ,16 ). In this paper we show that these Elav-like proteins also bind to the poly(A) tail of mRNA. This is mediated by the third RRM and may promote an interaction between AU-rich elements and the poly(A) tails of mRNA.

MATERIALS AND METHODS

Materials

HuR, HuD and deletion mutants of HuD were purified as GST fusion proteins as previously described (7 ,16 ). Poly(A)-Sepharose 4B and Sepharose 4B were from Pharmacia. BA 85 nitrocellulose filters were from Schleicher & Schuell. RNase T1 was obtained from Calbiochem. Poly(A), poly(G), poly(U) and poly(C) were from Sigma.

Preparation of RNA transcripts

RNA transcripts were synthesized from plasmid DNA using [[alpha]-32P]ATP or [32P]GTP and were gel purified before use (16 ). The specific activity of these transcripts is expressed as c.p.m./pmol nucleotide. The 3' myc and 3' myc(A)87 transcripts were derived from AflII and HindIII digests of pMycSD3, a gift of Dr Gary Brewer (24 ). The 3' myc(I) transcript was derived from a SspI digest of pMycSD3. The 3' myc(II) transcript was derived from an AflII digest of pMycSD3/[Delta]5. pMycSD3/[Delta]5 was created by deletion of an EcoRI-ApoI fragment from pMycSD3. The (A)87 transcript was derived from a HindIII digest of pSD3, also a gift of Dr Gary Brewer. Poly(A)600av was hydrolyzed with alkali and end-labeled using T4 kinase and [[gamma]-32P]ATP to a specific activity of 1 * 106 c.p.m./pmol ends and gel purified.

RNase T1 selection assay


Figure 1. The structure of the c-myc 3'-UTR and of the RNA transcripts used in this study. (A) Hex1 and Hex2 indicate the hexanucleotide poly(A) addition signals for the minor and major transcripts respectively. The major transcript is shown. AU1 and AU2 denote the HuR and HuD binding sites. The open squares indicate the Sp6 promoter elements. (B) The sequence of 3' myc(A)87. The shaded boxes indicate the binding sites for HuR and HuD. The Hex1 and Hex 2 poly(A) addition signals are indicated by the enclosed boxes of AAUAAA.


Figure 2. HuR binds to the AU-rich elements and poly(A) tail of 3' myc(A)87. (A) 32P-Labeled 3' myc(A)87 RNA (14 fmol, 1 * 105 c.p.m./pmol GMP) was incubated with the indicated concentrations of HuR and GST at 37oC for 10 min. After treating the reaction mixture with RNase T1 the reaction mixtures were filtered through nitrocellulose. RNA fragments bound to the nitrocellulose were extracted and electrophoresed in a 12% acrylamide-8 M urea gel. Lane 5 shows the T1 digest of the transcript prior to selection. (B) The indicated 32P-labeled RNAs (20 fmol, 1 * 104 c.p.m./pmol AMP) were incubated with HuR at 37oC for 10 min. In the first and second panels the concentration of HuR was 5, 10, 20, 50, 100 and 200 nM in lanes 1-6 respectively and the GST concentration was 200 nM. In the third panel the concentration of HuR was 20, 50, 100, 200, 500 and 1000 nM in lanes 1-6 respectively and the GST concentration was 1000 nM. The reaction mixtures were treated with RNase T1 and selected as described above. In each panel lane T shows the T1 digest of the transcript prior to selection. The amount loaded corresponds to 50% of the transcript used in the selection.

Reaction mixtures (0.02 ml) contained 50 mM Tris, pH 7.0, 150 mM NaCl, 0.25 mg/ml BSA, 0.25 mg/ml tRNA, radiolabeled RNA and purified HuR as indicated. After 10 min incubation at 37oC, RNase T1 (5 U) was added and the reaction continued for a further 10 min. The mixtures were diluted 1:6 with buffer F (20 mM Tris, pH 7.0, 150 mM NaCl) and filtered through nitrocellulose (BA 85; Schleicher & Schuell). After washing the nitrocellulose twice with buffer F, the bound RNA was eluted by phenol/chloroform extraction. The resultant RNA was mixed with formamide buffer, denatured at 65oC for 3 min and analyzed by 12% polyacrylamide-urea gel electrophoresis. The gel was fixed with 1:1:8 acetic acid:methanol:water, dried on DE-81 paper with a backing of gel drying paper and exposed to the XAR5 film at -70oC overnight.

Nitrocellulose filter binding assay

Reaction mixtures (0.02 ml) contained 50 mM Tris, pH 7.0, 150 mM NaCl, 0.25 mg/ml BSA, 2.5 [mu]g/ml tRNA, labeled RNA and purified protein as indicated. After 10 min incubation at 37oC the mixtures were diluted 1:6 with buffer F and filtered through nitrocellulose (BA85; Schleicher & Schuell). After washing the filter twice with buffer F, bound radioactivity was determined by Cerenkov counting. Each point is corrected for the amount of RNA bound in the absence of protein, which is usually <1% of the input.

RESULTS

The Elav-like proteins bind to AU-rich elements and to poly(A)

We have previously shown that the human Elav-like proteins bind specifically to the 3'-UTR of c-myc mRNA (7 ,16 ). Using RNase T1 selection analysis we discovered that the 3' myc mRNA contains two independent HuR binding sites that we have labeled AU1 and AU2 (see Fig. 1 ). The two sites were confirmed by RNase T1 selection analysis of transcripts 3' myc(I) and 3' myc(II) (Fig. 1 ). In view of the relationship betweeen mRNA decay and polyadenylation (25 ), we decided to examine interaction of the Elav-like proteins with a polyadenylated transcript. Using pMycSD3 and [32P]GTP we synthesized a c-myc transcript that contained a poly(A) tail of 87 nt. The HuR binding sites on this transcript were assayed by the RNase T1 selection assay (7 ). As expected, HuR bound specifically to the AU1 and AU2 sites. (Fig. 2 A, lanes 1-4). We were surprised, however, to observe that a larger band was also selected (Fig. 2 A, lanes 3 and 4). This band was the same size as the (A)87 fragment present in the total RNase T1 digest of the transcript [the (A)87 fragment is labeled by virtue of a G residue in the restriction site at the end of the template DNA]. Thus we concluded that HuR exhibits a poly(A) binding activity. This activity was intrinsic to the HuR protein, as no fragments were selected by high concentrations of GST. We next determined whether HuR could directly bind to poly(A). Using the pSD3 plasmid (see Fig. 1 ) we synthesized the (A)87 tail itself. To increase the sensitivity of the assay we labeled the transcripts with [32P]ATP. Figure 2 B shows that HuR bound to (A)87, even in the absence of the AU1 and AU2 sites. Thus HuR has an independent poly(A) binding activity. It is important to point out that HuR exhibits a significantly lower affinity with (A)87 than with the AU1 and AU2 elements.


Figure 3. Size selection analysis of poly(A) binding activity. (A) Poly(A) of 600 nt (average size) was cleaved with dilute alkali and labeled to yield a uniform size distribution from 24 to 800 nt. This population of poly(A) was then labeled at the 5'-end using ATP and T4 kinase. The 32P-labeled poly(A)24-800 (50 fmol, specific activity 1.1 * 106 c.p.m./pmol) was then incubated with GST or HuR (200 nM) at 37oC. After 10 min the protein-bound poly(A) was selected by absorption to nitrocellulose, eluted and electrophoresed in a 6% acrylamide-8 M urea gel. The labeled poly(A)24-800 is shown in lane P. The selected products after incubation with no protein, GST or HuR protein are shown in lanes 1-3 respectively. A marker digest of [Phi]X174 DNA is shown in lane M. A quantitative analysis (by scanning in a PhosphorImager) of lanes P and 3 is shown in (B) and (C) respectively.

Characterization of the poly(A) binding activity

Next we investigated the properties of the poly(A) binding activity in more detail. We were surprised to observe that HuR did not bind to poly(A)30. Thus we investigated whether HuR had a requirement for longer poly(A) tails. End-labeled poly(A) of uniform size distribution was prepared and incubated with HuR or GST. The bound poly(A) was selected by nitrocellulose filtration and analyzed by gel electrophoresis. Figure 3 shows that binding was first detectable with poly(A)80, was half-maximal with poly(A)180 and saturated at poly(A)300. This result was confirmed by purification of poly(A) of defined size and determination of their binding affinity. As in previous studies, the interactions between the Elav-like proteins and RNA were quantitated using a nitrocellulose filter binding assay (7 ,26 ). A low concentration of labeled RNA was incubated with increasing concentrations of HuR. The reaction mixtures were filtered through nitrocellulose and the bound radioactivity determined. As predicted from the selection analysis, virtually no complex was formed with poly(A)30 (Fig. 4 A). Increasing reactivity was observed with poly(A)100, poly(A)150 and poly(A)200, reaching saturation at poly(A)300. A plot of log[complex/free poly(A)] versus log(HuR concentration) revealed a straight line in each case (Fig. 4 A). This suggested a simple interaction with little cooperativity. The affinity of HuR for poly(A) is significantly less than that for the AU-rich elements. This is quantitatively shown in Figure 4 B. The apparent Kd for 3' myc is 4 nM, whereas the apparent Kd for poly(A)300 is 146 nM. As before, the intrinsic nature of these activities is indicated by the lack of reactivity with GST (Fig. 4 B). There is also little difference in binding between 3' myc and 3'myc(A)87 (Fig. 4 B). Thus the AU-rich element is the primary determinant of HuR binding. Next we examined the specificity of the poly(A) binding activity. We investigated whether the HuR-poly(A) complex could be displaced by other homopolymers. Figure 4 C shows that the HuR-poly(A)87 complex was displaced by poly(A)600 (50% displacement at 0.2 molar excess) and to a lesser extent by poly(G)600 (50% displacement at 0.8 molar excess). Neither poly(U)600 (50% displacement at 32 molar excess) nor poly(C)600 (no displacement at 1000-fold molar excess) significantly displaced the HuR-poly(A)87 complex.


Figure 4. (A) The affinity of HuR for poly(A)30, poly(A)100, poly(A)150, poly(A)200, poly(A)300 and poly(A)400. RNA-protein complex formation was assayed by nitrocellulose filtration. An aliquot of 14 fmol each RNA (specific activity 1.1 * 106 c.p.m./pmol) was incubated with the indicated concentration of HuR for 10 min at 37oC. (i) Plot of percentage RNA bound versus log HuR concentration. (ii) Plot of log complex/free RNA versus log HuR concentration. (B) The affinity of HuR for 3' myc, 3' myc(A)87 and (A)300. RNA-protein complex formation was assayed by nitrocellulose filtration. 32P-Labeled 3' myc(A)87, 3' myc (9 and 13 fmol, specific activity 1 * 104 c.p.m./pmol AMP) and (A)300 (14 fmol, specific activity 1.1 * 106 c.p.m./pmol) were incubated with the indicated concentration of HuR and GST for 10 min at 37oC. (C) Competition analysis of poly(A) binding activity. RNA binding was determined by nitrocellulose filtration. p(A)87 (20 fmol, specific activity 1 * 104 c.p.m./pmol) was incubated with 200 nM recombinant HuR and the indicated molar excess of poly(A)600, poly(G)600, poly(U)600 and poly(C)600. Eight femtomoles of p(A)87 were bound in the absence of competitor and set as 100%.

The poly(A) binding activity is resident in the third RNA binding domain

The most striking and unique structural feature of the Elav-like family of proteins is the presence and organization of the three putative RNA recognition motifs (Fig. 5 ). In each case the two tandemly arranged RNA recognition motifs are connected to the third RNA recognition motif by a highly basic segment that we have termed the `basic segment'. We have previously shown that the first and second RRMs of HuD are necessary and sufficient for binding to AU-rich elements (16 ). We now sought to establish which domains are involved in poly(A) binding activity. We used the mutant constructs derived from HuD (16 ; Fig. 5 ). HuD bound to the ARE and poly(A) tail of 3' c-myc(A)87 (Fig. 5 , lane 2). The first and second RNA binding domains (HuD I,II) did not bind to the poly(A) tail. Although HuD I,II does bind to AU-rich elements, it does so with significantly lower affinity (the apparent Kd for HuD is 16 nM, whereas the apparent Kd for HuD I,II is 125 nM) (16 ). Thus, at the concentration used here, little or no binding to the AU1 and AU2 elements was anticipated. In contrast, the third RNA binding domain (HuD III) bound avidly to the poly(A) tail (Fig. 5 , lane 4). Thus we concluded that the first and second RNA binding domains interact with AU-rich elements whereas the third RNA binding domain interacts with the poly(A) tail.


Figure 5. Analysis of RNA binding domains. (A) Structure of the mutant HuD derivatives. The residues of HuD contained in each construct are as follows: pGEX-HuD I,II, 2-216; pGEX-HuD III, 279-373. (B) The purified HuD derivatives (200 nM) were mixed with 32P-labeled 3' myc(A)87 (100 fmol, specific activity 5 * 103 c.p.m./pmol AMP). Following incubation at 37oC for 10 min the reaction mixtures were analyzed by the RNase T1 selection assay.

The Elav-like proteins bind simultaneously to the AU-rich element and the poly(A) tail of mRNA

Next we investigated whether the Elav-like proteins can contact both sites simultaneously. To examine this, we bound HuR to poly(A)-Sepharose beads (HuR/pA-S4B beads), removed unbound HuR by washing and then examined the ability of the beads to bind the AU-rich element. Figure 6 shows that the labeled 3' myc transcript bound to poly(A)-Sepharose beads preincubated with HuR (HuR/pA-S4B beads) but not to poly(A)-Sepharose beads preincubated with GST (GST/pA-S4B beads). The amount of transcript bound increased with increasing concentration of HuR. The labeled 3' myc transcript did not bind to Sepharose beads that were preincubated with HuR (HuR/S4B beads). This is a difficult experiment to perform since the off-rate of the HuR-poly(A) complex is fast. Thus the demonstration that 10% of the myc transcript can be simultaneously bound at saturating HuR concentration is significant. We concluded that the Elav-like proteins can form a bridge between the poly(A) tail and the AU-rich element.

DISCUSSION

The Elav-like proteins, HuD, HuC, Hel-N1 and HuR, stabilize specific mRNAs via an interaction with AU-rich elements in their 3'-UTR (7 ,14 -17 ,20 -22 ). In this paper we have shown that HuD and HuR proteins have an additional property, namely a novel poly(A) binding activity. Recently (after this paper was submitted for publication) the HuC protein was also shown to have poly(A) binding activity (27 ). Thus this activity is probably a feature of all Elav-like proteins. This activity may not have been detected in previous studies of Hel-N1 since poly(A) was used as a non-specific competitor (17 ,28 ). The properties of the Elav-like poly(A) binding activity are quite different from other poly(A) binding proteins. The cytoplasmic poly(A) binding protein (PABI) binds to (A)10 and the Kd does not significantly change with increased chain length (29 ). The nuclear poly(A) binding protein (PABII) binds to an oligo(A) tail >10-11 nt and remains associated [in a complex with poly(A) polymerase] until the tail is elongated to a length of 250 nt (30 ). Thus these activities have a minimal binding site of 10-15 nt, whereas the Elav-like proteins prefer polymers >70 nt. The affinity of the Elav-like proteins for poly(A) is relatively low [an apparent Kd of 146 nM for (A)300] compared with a Kd of 5 nM for PABI (29 ,31 ) and a Kd of 2 nM for PABII (32 ). The Elav-like poly(A) binding activity is, however, similar to both PABI and PABII in that its activity is displaced by a molar excess of poly(A) and poly(G) but not efficiently by poly(U) or poly(C). The observation that the Elav-like proteins preferentially bind to long polymers of poly(A) is surprising. Typically, proteins exhibit increased binding to polymers as a result of protein-protein interactions. This does not appear to be the case with the Elav-like proteins. There is no obvious cooperativity in their interaction with either short or long chain poly(A). Thus the possibility remains that the third RNA binding domain recognizes a secondary or tertiary structure that is only evident in long chain poly(A).


Figure 6. HuR binds to poly(A) and ARE simultaneously. Sepharose 4B and poly(A)-Sepharose 4B beads were preincubated with the indicated concentration of HuR in a 50 [mu]l reaction containing 50 mM Tris-HCl, pH 7.0, 150 mM NaCl, 2.5 [mu]g tRNA/ml, 250 [mu]g BSA/ml and 0.01% NP-40. The mixture was shaken at room temperature for 1 h. The beads were spun down and the supernatant removed. The beads were washed with 200 [mu]l wash buffer (50 mM Tris-HCl, pH 7.0, 150 mM NaCl, 0.01% NP-40). The washed beads were then resuspended in 50 [mu]l containing 50 mM Tris-HCl, pH 7.0, 150 mM NaCl, 2.5 [mu]g tRNA/ml, 250 [mu]g BSA/ml and 0.01% NP-40 and 3' c-myc RNA transcript (50 fmol, specific activity 5.0 * 103 c.p.m./pmol AMP). The mixture was incubated at room temperature for 15 min with shaking. The beads were washed with 3 ml wash buffer (500 [mu]l each wash). Bound radioactivity was determined by Cerenkov counting.

The Elav-like proteins contain three RNA binding domains. Previous studies have shown that the third RNA binding domain is not required for AU-rich element binding (16 ). This domain is crucial, however, since mutation of Gly426 to glutamic acid in Drosophila Elav leads to a temperature-sensitive phenotype (33 ). We have shown here that the third RNA binding domain contains the poly(A) binding activity. The bifunctionality of the Elav-like proteins is not surprising. The first and second RNA binding domains are much more closely related in sequence to the corresponding domains among different organisms than to the third RNA binding domain (2 ,5 -7 ,13 ). This supports the notion that they have different functions.

The demonstration of a poly(A) binding activity resident in the third RNA binding domain of the Elav-like proteins has important implications for understanding their mechanism of action. Our current model is that binding of the Elav-like protein inhibits the action of a specific endonuclease that recognizes the AU-rich element. It is possible that effective inhibition of the endonuclease may require association of the Elav-like proteins with both the AU-rich element and the poly(A) tail. This would ensure that deadenylated message would not be stabilized. Alternatively, the Elav-like proteins may sequester the poly(A) tail of a target mRNA, with a consequent inhibitory effect on a poly(A) exonuclease activity. The poly(A) mRNA-protein complex described here and the recent development of an in vitro system that recapitulates the regulated turnover of mRNA (21 ) may provide a way to answer these questions.

ACKNOWLEDGEMENTS

We thank Barry Nevins for his patience in preparing the manuscript. This work was supported by the NIH (NS29682), The Byrne Fund and an NCI core grant (P30-CA08748).

REFERENCES

1 Robinow,S., Campos,A.R., Yao,K.M. and White,K. (1988) Science, 242, 1570-1572. MEDLINE Abstract

2 Abe,R., Uyeno,Y., Yamamoto,K. and Sakamoto,H. (1994) DNA Res., 1, 175-180. MEDLINE Abstract

3 King,P.H., Levine,T.D., Fremeau,R.T. and Keene,J.D. (1994) J. Neurosci., 14, 1943-1952. MEDLINE Abstract

4 Sakai,K., Gofuku,M., Kitagawa,Y., Ogasawara,T., Hirose,G., Yamazaki,M., Koh,C., Yanagisawa,N. and Steinman,L. (1994) Biochem. Biophys. Res. Commun., 199, 1200-1208. MEDLINE Abstract

5 Szabo,A., Dalmau,J., Manley,G., Rosenfeld,M.R., Wong,E., Henson,J., Posner,J.B. and Furneaux,H.M. (1991) Cell, 67, 325-333. MEDLINE Abstract

6 Perron,M., Theodore,L. and Wegnez,M. (1995) Mech. Dev., 51, 235-249. MEDLINE Abstract

7 Ma,W.-J., Cheng,S., Wright,A., Campbell,C. and Furneaux,H.M. (1996) J. Biol. Chem., 271, 8144-8151.

8 Good,P.J. (1995) Proc. Natl. Acad. Sci. USA, 92, 4557-4561. MEDLINE Abstract

9 Marusich,H.M., Furneaux,H.M., Henion,P. and Weston,J.A. (1994) J. Neurobiol., 25, 143-155.

10 Barami,K., Iversen,K., Furneaux,H. and Goldman,S. (1995) J. Neurobiol., 28, 82-101. MEDLINE Abstract

11 Graus,F., Cordon-Cardo,C. and Posner,J.B. (1985) Neurology, 35, 538-543. MEDLINE Abstract

12 Dalmau,J., Furneaux,H.M., Cordon-Cardo,C. and Posner,J.B. (1992) Am. J. Pathol., 141, 1-6.

13 Liu,J., Dalmau,J., Szabo,A., Rosenfeld,M., Huber,J. and Furneaux,H. (1995) Neurology, 45, 544-550. MEDLINE Abstract

14 Chung,S., Perrone-Bizzozero,N., Kohn,D.T. and Furneaux,H. (1997) J. Biol. Chem., 272, 6593-6598. MEDLINE Abstract

15 Levine,T.D., Gao,F., King,P.H., Andrews,L.C. and Keene,J.D. (1993) Mol. Cell. Biol., 13, 3494-3504. MEDLINE Abstract

16 Chung,S., Jiang,L., Cheng,S. and Furneaux,H.M. (1996) J. Biol. Chem., 271, 11518-11524. MEDLINE Abstract

17 Gao,F.B., Carson,C., Levine,T. and Keene,J. (1994) Proc. Natl. Acad. Sci. USA, 91, 11207-11211. MEDLINE Abstract

18 Shaw,G. and Kamen,R. (1986) Cell, 46, 659-667. MEDLINE Abstract

19 Ross,J. (1995) Microbiol. Rev., 59, 423-450. MEDLINE Abstract

20 Aranda-Abreu,G., Behar,L., Chung,S., Furneaux,H. and Ginzburg,I. (1997) J. Biol. Chem., submitted for publication.

21 Levy,N.S., Chung,S., Furneaux,H.M. and Levy,A.P. (1997) J. Biol. Chem., submitted for publication.

22 Jain,R.G., Andrews,L.G., McGowan,K.M., Pekala,P.H. and Keene,J.D. (1997) Mol. Cell. Biol., 17, 954-962. MEDLINE Abstract

23 Myer,V.E., Fan,X.C. and Steitz,J.A. (1997) EMBO J., 16, 2130-2139. MEDLINE Abstract

24 Brewer,G. (1991) Mol. Cell. Biol., 11, 2460-2466. MEDLINE Abstract

25 Wilson,T. and Treisman,R. (1988) Nature, 336, 396-399. MEDLINE Abstract

26 Carey,J., Cameron,V., de Haseth,P.L. and Uhlenbeck,O.C. (1983) Biochemistry, 22, 2601-2610. MEDLINE Abstract

27 Abe,R., Sakashita,E., Yamamoto,K. and Sakamoto,H. (1996) Nucleic Acids Res., 24, 4895-4901. MEDLINE Abstract

28 Levine,T.D., Gao,F., King,P.H., Andrews,L.C. and Keene,J.D. (1993) Mol. Cell. Biol., 13, 3494-3504. MEDLINE Abstract

29 Sachs,A.B., Davis,R.W. and Kornberg,R.D. (1987) Mol. Cell. Biol., 7, 3268-3276. MEDLINE Abstract

30 Wahle,E., Martin,G., Schiltz,E. and Keller,W. (1991) EMBO J., 10, 4251-4257. MEDLINE Abstract

31 Gorlach,M., Burd,C.G. and Dreyfuss,G. (1994) Exp. Cell. Res., 211, 400-407. MEDLINE Abstract

32 Wahle,E., Lustig,A., Jeno,P. and Maurer,P. (1993) J. Biol. Chem., 268, 2937-45. MEDLINE Abstract

33 Samson,M.L., Lisbin,M.J. and White,K. (1995) Genetics, 141, 1101-1111. MEDLINE Abstract

* To whom correspondence should be addressed. Tel: +1 212 639 8701; Fax: +1 212 639 2861; Email: h-furneaux@ski.mskcc.org
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Antagonistic Functions of Tetradecanoyl Phorbol Acetate-Inducible-Sequence 11b and HuR in the Hormonal Regulation of Vascular Endothelial Growth Factor Messenger Ribonucleic Acid Stability by Adrenocorticotropin
Mol. Endocrinol., April 1, 2006; 20(4): 916 - 930.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
T. Fujiwara, Y. Mori, D. L. Chu, Y. Koyama, S. Miyata, H. Tanaka, K. Yachi, T. Kubo, H. Yoshikawa, and M. Tohyama
CARM1 Regulates Proliferation of PC12 Cells by Methylating HuD.
Mol. Cell. Biol., March 1, 2006; 26(6): 2273 - 2285.
[Abstract] [Full Text] [PDF]

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 Nucleic Acids ResHome page
C. Barreau, L. Paillard, and H. B. Osborne
AU-rich elements and associated factors: are there unifying principles?
Nucleic Acids Res., January 3, 2006; 33(22): 7138 - 7150.
[Abstract] [Full Text] [PDF]

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 Nucleic Acids ResHome page
C. D. Borgeson and M.-L. Samson
Shared RNA-binding sites for interacting members of the Drosophila ELAV family of neuronal proteins
Nucleic Acids Res., November 10, 2005; 33(19): 6372 - 6383.
[Abstract] [Full Text] [PDF]

Home page
 J. Leukoc. Biol.Home page
M. T. Pritchard, Z. Li, and E. A. Repasky
Nitric oxide production is regulated by fever-range thermal stimulation of murine macrophages
J. Leukoc. Biol., September 1, 2005; 78(3): 630 - 638.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
W. Wang, X. Yang, T. Kawai, I. L. de Silanes, K. Mazan-Mamczarz, P. Chen, Y. M. Chook, C. Quensel, M. Kohler, and M. Gorospe
AMP-activated Protein Kinase-regulated Phosphorylation and Acetylation of Importin {alpha}1: INVOLVEMENT IN THE NUCLEAR IMPORT OF RNA-BINDING PROTEIN HuR
J. Biol. Chem., November 12, 2004; 279(46): 48376 - 48388.
[Abstract] [Full Text] [PDF]

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 Proc. Natl. Acad. Sci. USAHome page
A. Pascale, P. A. Gusev, M. Amadio, T. Dottorini, S. Govoni, D. L. Alkon, and A. Quattrone
Increase of the RNA-binding protein HuD and posttranscriptional up-regulation of the GAP-43 gene during spatial memory
PNAS, February 3, 2004; 101(5): 1217 - 1222.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
L.-M. Buu, L.-T. Jang, and F.-J. S. Lee
The Yeast RNA-binding Protein Rbp1p Modifies the Stability of Mitochondrial Porin mRNA
J. Biol. Chem., January 2, 2004; 279(1): 453 - 462.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
K. Aoki, K. Matsumoto, and M. Tsujimoto
Xenopus Cold-inducible RNA-binding Protein 2 Interacts with ElrA, the Xenopus Homolog of HuR, and Inhibits Deadenylation of Specific mRNAs
J. Biol. Chem., November 28, 2003; 278(48): 48491 - 48497.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
S. Park-Lee, S. Kim, and I. A. Laird-Offringa
Characterization of the Interaction between Neuronal RNA-binding Protein HuD and AU-rich RNA
J. Biol. Chem., October 10, 2003; 278(41): 39801 - 39808.
[Abstract] [Full Text] [PDF]

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 Mol. Cell. Biol.Home page
A. Figueroa, A. Cuadrado, J. Fan, U. Atasoy, G. E. Muscat, P. Munoz-Canoves, M. Gorospe, and A. Munoz
Role of HuR in Skeletal Myogenesis through Coordinate Regulation of Muscle Differentiation Genes
Mol. Cell. Biol., July 15, 2003; 23(14): 4991 - 5004.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
S. Sengupta, B.-C. Jang, M.-T. Wu, J.-H. Paik, H. Furneaux, and T. Hla
The RNA-binding Protein HuR Regulates the Expression of Cyclooxygenase-2
J. Biol. Chem., June 27, 2003; 278(27): 25227 - 25233.
[Abstract] [Full Text] [PDF]

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 Mol. Biol. CellHome page
R. Shao, Z. Shi, P. J. Gotwals, V. E. Koteliansky, J. George, and D. C. Rockey
Cell and Molecular Regulation of Endothelin-1 Production during Hepatic Wound Healing
Mol. Biol. Cell, June 1, 2003; 14(6): 2327 - 2341.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
M. T. Worthington, J. W. Pelo, M. A. Sachedina, J. L. Applegate, K. O. Arseneau, and T. T. Pizarro
RNA Binding Properties of the AU-rich Element-binding Recombinant Nup475/TIS11/Tristetraprolin Protein
J. Biol. Chem., December 6, 2002; 277(50): 48558 - 48564.
[Abstract] [Full Text] [PDF]

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 Mol. Cell. Biol.Home page
C.-Y. A. Chen, N. Xu, and A.-B. Shyu
Highly Selective Actions of HuR in Antagonizing AU-Rich Element-Mediated mRNA Destabilization
Mol. Cell. Biol., October 15, 2002; 22(20): 7268 - 7278.
[Abstract] [Full Text] [PDF]

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 Nucleic Acids ResHome page
K. Kasashima, E. Sakashita, K. Saito, and H. Sakamoto
Complex formation of the neuron-specific ELAV-like Hu RNA-binding proteins
Nucleic Acids Res., October 15, 2002; 30(20): 4519 - 4526.
[Abstract] [Full Text] [PDF]

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 Am. J. Physiol. Heart Circ. Physiol.Home page
K. Tang, E. C. Breen, and P. D. Wagner
Hu protein R-mediated posttranscriptional regulation of VEGF expression in rat gastrocnemius muscle
Am J Physiol Heart Circ Physiol, October 1, 2002; 283(4): H1497 - H1504.
[Abstract] [Full Text] [PDF]

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 Nucleic Acids ResHome page
J. Milone, J. Wilusz, and V. Bellofatto
Identification of mRNA decapping activities and an ARE-regulated 3' to 5' exonuclease activity in trypanosome extracts
Nucleic Acids Res., September 15, 2002; 30(18): 4040 - 4050.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
A. C. Beckel-Mitchener, A. Miera, R. Keller, and N. I. Perrone-Bizzozero
Poly(A) Tail Length-dependent Stabilization of GAP-43 mRNA by the RNA-binding Protein HuD
J. Biol. Chem., July 26, 2002; 277(31): 27996 - 28002.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
B. B. Yeap, D. C. Voon, J. P. Vivian, R. K. McCulloch, A. M. Thomson, K. M. Giles, M. F. Czyzyk-Krzeska, H. Furneaux, M. C. J. Wilce, J. A. Wilce, et al.
Novel Binding of HuR and Poly(C)-binding Protein to a Conserved UC-rich Motif within the 3'-Untranslated Region of the Androgen Receptor Messenger RNA
J. Biol. Chem., July 19, 2002; 277(30): 27183 - 27192.
[Abstract] [Full Text] [PDF]

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 Mol. Cell. Biol.Home page
W. Wang, J. Fan, X. Yang, S. Furer-Galban, I. Lopez de Silanes, C. von Kobbe, J. Guo, S. N. Georas, F. Foufelle, D. G. Hardie, et al.
AMP-Activated Kinase Regulates Cytoplasmic HuR
Mol. Cell. Biol., May 15, 2002; 22(10): 3425 - 3436.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
I. Goldberg-Cohen, H. Furneauxb, and A. P. Levy
A 40-bp RNA Element That Mediates Stabilization of Vascular Endothelial Growth Factor mRNA by HuR
J. Biol. Chem., April 12, 2002; 277(16): 13635 - 13640.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
R. S. Phillips, S. B. V. Ramos, and P. J. Blackshear
Members of the Tristetraprolin Family of Tandem CCCH Zinc Finger Proteins Exhibit CRM1-dependent Nucleocytoplasmic Shuttling
J. Biol. Chem., March 22, 2002; 277(13): 11606 - 11613.
[Abstract] [Full Text] [PDF]

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 J. Cell Sci.Home page
S. Aronov, G. Aranda, L. Behar, and I. Ginzburg
Visualization of translated tau protein in the axons of neuronal P19 cells and characterization of tau RNP granules
J. Cell Sci., January 10, 2002; 115(19): 3817 - 3827.
[Abstract] [Full Text] [PDF]

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 ScienceHome page
I.-E. Gallouzi and J. A. Steitz
Delineation of mRNA Export Pathways by the Use of Cell-Permeable Peptides
Science, November 30, 2001; 294(5548): 1895 - 1901.
[Abstract] [Full Text] [PDF]

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 Proc. Natl. Acad. Sci. USAHome page
A. Quattrone, A. Pascale, X. Nogues, W. Zhao, P. Gusev, A. Pacini, and D. L. Alkon
Posttranscriptional regulation of gene expression in learning by the neuronal ELAV-like mRNA-stabilizing proteins
PNAS, September 25, 2001; 98(20): 11668 - 11673.
[Abstract] [Full Text] [PDF]

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 Genes Dev.Home page
G. K. Voeltz, J. Ongkasuwan, N. Standart, and J. A. Steitz
A novel embryonic poly(A) binding protein, ePAB, regulates mRNA deadenylation in Xenopus egg extracts
Genes & Dev., March 15, 2001; 15(6): 774 - 788.
[Abstract] [Full Text]

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 Proc. Natl. Acad. Sci. USAHome page
S. A. Tenenbaum, C. C. Carson, P. J. Lager, and J. D. Keene
Identifying mRNA subsets in messenger ribonucleoprotein complexes by using cDNA arrays
PNAS, December 19, 2000; 97(26): 14085 - 14090.
[Abstract] [Full Text] [PDF]

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 J. Cell Biol.Home page
C. M. Brennan, I.-E. Gallouzi, and J. A. Steitz
Protein Ligands to HuR Modulate Its Interaction with Target mRNAs In Vivo
J. Cell Biol., September 25, 2000; 151(1): 1 - 14.
[Abstract] [Full Text] [PDF]

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 Nucleic Acids ResHome page
Z. Zhao, F.-C. Chang, and H. M. Furneaux
The identification of an endonuclease that cleaves within an HuR binding site in mRNA
Nucleic Acids Res., July 15, 2000; 28(14): 2695 - 2701.
[Abstract] [Full Text] [PDF]

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 Mol. Cell. Biol.Home page
S. Park, D. G. Myszka, M. Yu, S. J. Littler, and I. A. Laird-Offringa
HuD RNA Recognition Motifs Play Distinct Roles in the Formation of a Stable Complex with AU-Rich RNA
Mol. Cell. Biol., July 1, 2000; 20(13): 4765 - 4772.
[Abstract] [Full Text]

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 Proc. Natl. Acad. Sci. USAHome page
I.-E. Gallouzi, C. M. Brennan, M. G. Stenberg, M. S. Swanson, A. Eversole, N. Maizels, and J. A. Steitz
HuR binding to cytoplasmic mRNA is perturbed by heat shock
PNAS, March 28, 2000; 97(7): 3073 - 3078.
[Abstract] [Full Text] [PDF]

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 Mol. Cell. Biol.Home page
W. Wang, H. Furneaux, H. Cheng, M. C. Caldwell, D. Hutter, Y. Liu, N. Holbrook, and M. Gorospe
HuR Regulates p21 mRNA Stabilization by UV Light
Mol. Cell. Biol., February 1, 2000; 20(3): 760 - 769.
[Abstract]