Nucleic Acids Research, 2000, Vol. 28, No. 5 1139-1144
© 2000 Oxford University Press
Characterization of the E.coli poly(A) polymerase: nucleotide specificity, RNA-binding affinities and RNA structure dependence
Department of Biology, Technion–Israel Institute of Technology, Haifa 32000, Israel
Polyadenylation of RNA molecules in bacteria and chloroplasts has been implicated as part of the RNA degradation pathway. The polyadenylation reaction is performed in Escherichia coli mainly by the enzyme poly(A) polymerase I (PAP I). In order to understand the molecular mechanism of RNA polyadenylation in bacteria, we characterized the biochemical properties of this reaction in vitro using the purified enzyme. Unlike the PAP from yeast nucleus, which is specific for ATP, E.coli PAP I can use all four nucleotide triphosphates as substrates for addition of long ribohomopolymers to RNA. PAP I displays a high binding activity to poly(U), poly(C) and poly(A) ribohomopolymers, but not to poly(G). The 3'-ends of most of the mRNA molecules in bacteria are characterized by a stem–loop structure. We show here that in vitro PAP I activity is inhibited by a stem–loop structure. A tail of two to six nucleotides located 3' to the stem–loop structure is sufficient to overcome this inhibition. These results suggest that the stem–loop structure located in most of the mRNA 3'-ends may function as an inhibitor of polyadenylation and degradation of the corresponding RNA molecule. However, RNA 3'-ends produced by endonucleolytic cleavage by RNase E in single-strand regions of mRNA molecules may serve as efficient substrates for polyadenylation that direct these molecules for rapid exonucleolytic degradation.
* To whom correspondence should be addressed. Tel: +972 4 8293171; Fax: +972 4 8225153; Email: gadis@tx.technion.ac.il
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  A. Just, F. Butter, M. Trenkmann, T. Heitkam, M. Morl, and H. Betat A comparative analysis of two conserved motifs in bacterial poly(A) polymerase and CCA-adding enzyme Nucleic Acids Res., August 5, 2008; (2008) gkn494v1. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. van den Hout, S. Hage, C. Dekker, and N. H. Dekker End-joining long nucleic acid polymers Nucleic Acids Res., July 25, 2008; (2008) gkn442v1. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. Martin and W. Keller RNA-specific ribonucleotidyl transferases RNA, November 1, 2007; 13(11): 1834 - 1849. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. D. Cho, C. L. M. J. Verlinde, and A. M. Weiner Reengineering CCA-adding enzymes to function as (U,G)- or dCdCdA-adding enzymes or poly(C,A) and poly(U,G) polymerases PNAS, January 2, 2007; 104(1): 54 - 59. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Bralley, S. A. Chang, and G. H. Jones A Phylogeny of Bacterial RNA Nucleotidyltransferases: Bacillus halodurans Contains Two tRNA Nucleotidyltransferases J. Bacteriol., September 1, 2005; 187(17): 5927 - 5936. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Hema, K. Gopinath, and C. Kao Repair of the tRNA-Like CCA Sequence in a Multipartite Positive-Strand RNA Virus J. Virol., February 1, 2005; 79(3): 1417 - 1427. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Sohlberg, J. Huang, and S. N. Cohen The Streptomyces coelicolor Polynucleotide Phosphorylase Homologue, and Not the Putative Poly(A) Polymerase, Can Polyadenylate RNA J. Bacteriol., December 15, 2003; 185(24): 7273 - 7278. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Yehudai-Resheff, V. Portnoy, S. Yogev, N. Adir, and G. Schuster Domain Analysis of the Chloroplast Polynucleotide Phosphorylase Reveals Discrete Functions in RNA Degradation, Polyadenylation, and Sequence Homology with Exosome Proteins PLANT CELL, September 1, 2003; 15(9): 2003 - 2019. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Le Derout, M. Folichon, F. Briani, G. Deho, P. Regnier, and E. Hajnsdorf Hfq affects the length and the frequency of short oligo(A) tails at the 3' end of Escherichia coli rpsO mRNAs Nucleic Acids Res., July 15, 2003; 31(14): 4017 - 4023. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Yehudai-Resheff, M. Hirsh, and G. Schuster Polynucleotide Phosphorylase Functions as Both an Exonuclease and a Poly(A) Polymerase in Spinach Chloroplasts Mol. Cell. Biol., August 15, 2001; 21(16): 5408 - 5416. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Kuhn, U. Tengler, and S. Binder Transcript Lifetime Is Balanced between Stabilizing Stem-Loop Structures and Degradation-Promoting Polyadenylation in Plant Mitochondria Mol. Cell. Biol., February 1, 2001; 21(3): 731 - 742. [Abstract] [Full Text]
|
|
|
|
|
|
B. K. Mohanty and S. R. Kushner Polynucleotide phosphorylase functions both as a 3' right-arrow 5' exonuclease and a poly(A) polymerase in Escherichiacoli PNAS, October 12, 2000; (2000) 220295997. [Abstract] [Full Text]
|
|
|
|
|
|
B. K. Mohanty and S. R. Kushner Polynucleotide phosphorylase functions both as a 3' right-arrow 5' exonuclease and a poly(A) polymerase in Escherichiacoli PNAS, October 24, 2000; 97(22): 11966 - 11971. [Abstract] [Full Text] [PDF]
|
|