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Author Notes:

*To whom correspondence should be addressed. Tel: +1 404 727 3924; Fax: 404 727 3949; Email: jfridov@emory.edu

We gratefully acknowledge Brad Bowzard, who helped enormously with the FPLC work, Kim Openo, who helped enormously with the complementation assay, and all other members of the Fridovich-Keil laboratory for their many helpful discussions and contributions.

We also thank Ryan Mills for his help with the SMART and Pfam sequence analysis systems, and Ms. Cheryl Strauss for expert editing of the manuscript.

Conflict of interest statement. None declared.

Subject:

Research Funding:

This work was supported in part by funds from the National Science Foundation (to JLFK), and in part by funds from the Emory University Research Committee (to JLFK).

MB was also supported in part by funds from the National Institutes of Health Training Grant T32 GM-08490 (PI J. Lucchesi).

Funding to pay the Open Access publication charge was provided by Emory University.

Functional overlap between conserved and diverged KH domains in Saccharomyces cerevisiae SCP160

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Journal Title:

Nucleic Acids Research

Volume:

Volume 35, Number 4

Publisher:

, Pages 1108-1118

Type of Work:

Article | Final Publisher PDF

Abstract:

The K homology (KH) domain is a remarkably versatile and highly conserved RNA-binding motif. Classical KH domains include a characteristic pattern of hydrophobic residues, a Gly-X-X-Gly (GXXG) segment, and a variable loop. KH domains typically occur in clusters, with some retaining their GXXG sequence (conserved), while others do not (diverged). As a first step towards addressing whether GXXG is essential for KH-domain function, we explored the roles of conserved and diverged KH domains in Scp160p, a multiple-KH-domain-containing protein in Saccharomyces cerevisiae. We specifically wanted to know (1) whether diverged KH domains were essential for Scp160p function, and (2) whether diverged KH domains could functionally replace conserved KH domains. To address these questions, we deleted and/or interchanged conserved and diverged KH domains of Scp160p and expressed the mutated alleles in yeast. Our results demonstrated that the answer to each question was yes. Both conserved and diverged KH domains are essential for Scp160p function, and diverged KH domains can function in place of conserved KH domains. These findings challenge the prevailing notions about the requisite features of a KH domain and raise the possibility that there may be more functional KH domains in the proteome than previously appreciated.

Copyright information:

© 2007 The Author(s).

This is an Open Access work distributed under the terms of the Creative Commons Attribution-NonCommercial 2.0 Generic License (http://creativecommons.org/licenses/by-nc/2.0/).

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