Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (429K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (22)
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Martin, F.
Right arrow Articles by Schümperli, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Martin, F.
Right arrow Articles by Schümperli, D.
Related Collections
Right arrow Protein-nucleic acid interaction
Social Bookmarking
 Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2000, Vol. 28, No. 7 1594-1603
© 2000 Oxford University Press

Positive and negative mutant selection in the human histone hairpin-binding protein using the yeast three-hybrid system

Franck Martin1,2, Fabrice Michel1, Daniel Zenklusen1, Berndt Müller1 and Daniel Schümperli1,*

1Abteilung für Entwicklungsbiologie, Zoologisches Institut der Universität Bern, Baltzerstrasse 4, 3012 Bern, Switzerland and 2Institut de Biologie Moléculaire et Cellulaire du CNRS, UPR 9002, 15 rue René Descartes, F-67084 Strasbourg Cedex, France

We have used the yeast three-hybrid system in a positive selection for mutants of the human histone hairpin-binding protein (HBP) capable of interacting with non-canonical hairpins and in a negative selection for loss-of-binding mutants. Interestingly, all mutations from the positive selection are located in the N- and C-terminal regions flanking a minimal RNA-binding domain (RBD) previously defined between amino acids 126 and 198. Further, in vitro binding studies demonstrate that the RBD, which shows no obvious similarity to other RNA-binding motifs, has a relaxed sequence specificity compared to full-length HBP, allowing it to bind to mutant hairpin RNAs not normally found in histone genes. These findings indicate that the sequences flanking the RBD are important for restricting binding to the highly conserved histone hairpin structure. Among the loss-of-binding mutations, about half are nonsense mutations distributed throughout the N-terminal part and the RBD whereas the other half are missense mutations restricted to the RBD. Whereas the nonsense mutations permit a more precise definition of the C-terminal border of the RBD, the missense mutations identify critical residues for RNA binding within the RBD.

* To whom correspondence should be addressed. Tel: +41 31 631 4675; Fax: +41 31 631 4616; Email: daniel.schuemperli@zoi.unibe.ch Present addresses: Daniel Zenklusen, Institut de Microbiologie, Université de Lausanne, Rue du Bugnon 44, 1011 Lausanne, Switzerland Berndt Müller, Department of Molecular and Cell Biology, Institute of Medical Sciences, University of Aberdeen, Aberdeen AB25 2ZD, UK


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
Nucleic Acids ResHome page
S. Jaeger, F. Martin, J. Rudinger-Thirion, R. Giege, and G. Eriani
Binding of human SLBP on the 3'-UTR of histone precursor H4-12 mRNA induces structural rearrangements that enable U7 snRNA anchoring
Nucleic Acids Res., October 18, 2006; 34(17): 4987 - 4995.
[Abstract] [Full Text] [PDF]


Home page
J. Cell Sci.Home page
X. Zhao, S. McKillop-Smith, and B. Muller
The human histone gene expression regulator HBP/SLBP is required for histone and DNA synthesis, cell cycle progression and cell proliferation in mitotic cells
J. Cell Sci., December 1, 2004; 117(25): 6043 - 6051.
[Abstract] [Full Text] [PDF]


Home page
Nucleic Acids ResHome page
Y.-G. Zheng, H. Wei, C. Ling, F. Martin, G. Eriani, and E.-D. Wang
Two distinct domains of the {beta} subunit of Aquifex aeolicus leucyl-tRNA synthetase are involved in tRNA binding as revealed by a three-hybrid selection
Nucleic Acids Res., June 18, 2004; 32(11): 3294 - 3303.
[Abstract] [Full Text] [PDF]


Home page
J. Biol. Chem.Home page
R. Pudi, S. Abhiman, N. Srinivasan, and S. Das
Hepatitis C Virus Internal Ribosome Entry Site-mediated Translation Is Stimulated by Specific Interaction of Independent Regions of Human La Autoantigen
J. Biol. Chem., March 28, 2003; 278(14): 12231 - 12240.
[Abstract] [Full Text] [PDF]


Home page
Mol. Cell. Biol.Home page
R. Sanchez and W. F. Marzluff
The Stem-Loop Binding Protein Is Required for Efficient Translation of Histone mRNA In Vivo and In Vitro
Mol. Cell. Biol., October 15, 2002; 22(20): 7093 - 7104.
[Abstract] [Full Text] [PDF]


Home page
Genes Dev.Home page
Z. Dominski, J. A. Erkmann, X. Yang, R. Sanchez, and W. F. Marzluff
A novel zinc finger protein is associated with U7 snRNP and interacts with the stem-loop binding protein in the histone pre-mRNP to stimulate 3'-end processing
Genes & Dev., January 1, 2002; 16(1): 58 - 71.
[Abstract] [Full Text] [PDF]


Home page
Mol. Cell. Biol.Home page
Z. Dominski, J. A. Erkmann, J. A. Greenland, and W. F. Marzluff
Mutations in the RNA Binding Domain of Stem-Loop Binding Protein Define Separable Requirements for RNA Binding and for Histone Pre-mRNA Processing
Mol. Cell. Biol., March 15, 2001; 21(6): 2008 - 2017.
[Abstract] [Full Text]



Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.