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RNA. 2008 November; 14(11): 2333–2347.
doi: 10.1261/rna.1016508.
PMCID: PMC2578860
Distinct roles for Khd1p in the localization and expression of bud-localized mRNAs in yeast
Yuko Hasegawa,1,2 Kenji Irie,1,2 and André P. Gerber3
1Department of Molecular Cell Biology, Graduate School of Comprehensive Human Sciences, University of Tsukuba, 305-8575 Tsukuba, Japan
2Institute of Basic Medical Sciences, University of Tsukuba, 305-8575 Tsukuba, Japan
3Institute of Pharmaceutical Sciences, ETH Zurich, 8093 Zurich, Switzerland
Reprint requests to: Kenji Irie, Department of Molecular Cell Biology, Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennoudai, Tsukuba, 305-8575, Japan; e-mail: kirie/at/md.tsukuba.ac.jp; fax: 81-29-853-3066.
Received January 25, 2008; Accepted August 4, 2008.
Abstract
The RNA-binding protein Khd1p (KH-domain protein 1) is required for efficient localization of ASH1 mRNA to the bud-tip, probably acting as a translational repressor during mRNA transport in yeast. Here, we have systematically examined Khd1p mRNA targets and colocalization with known bud-tip-localized mRNAs in vivo. Affinity purification and DNA microarray analysis of Khd1p-associated mRNAs revealed hundreds of potential mRNAs targets, many of them encoding membrane-associated proteins. The putative targets include the messages for MID2, MTL1, WSC2, SRL1, EGT2, CLB2, ASH1, and Khd1p colocalizes with these mRNAs at the bud-tip. The combination of bioinformatics, RNA localization, and in vitro RNA-binding assays revealed that Khd1p binds to CNN repeats in coding regions of mRNA targets. Among the proteins encoded by previously known bud-tip-localized mRNAs, only Mtl1p levels were decreased in khd1Δ mutant cells, whereas Ash1p and Srl1p were reduced in cells overexpressing KHD1. Hence, Khd1p differentially affects gene expression possibly due to combinatorial arrangement with additional factors reflecting the redundant structure of post-transcriptional regulatory systems.
Keywords: mRNA localization, RNA-binding protein, KHD1, translation, mRNA stability, yeast