Distinct roles for Khd1p in the localization and expression of bud-localized mRNAs in yeast

被引:54
作者
Hasegawa, Yuko [1 ,2 ]
Irie, Kenji [1 ,2 ]
Gerber, Andre P. [3 ]
机构
[1] Univ Tsukuba, Dept Mol Cell Biol, Grad Sch Comprehens Human Sci, Tsukuba, Ibaraki 3058575, Japan
[2] Univ Tsukuba, Inst Basic Med Sci, Tsukuba, Ibaraki 3058575, Japan
[3] Swiss Fed Inst Technol, Inst Pharmaceut Sci, CH-8093 Zurich, Switzerland
关键词
mRNA localization; RNA-binding protein; KHD1; translation; mRNA stability; yeast;
D O I
10.1261/rna.1016508
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 khd1 Delta 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.
引用
收藏
页码:2333 / 2347
页数:15
相关论文
共 75 条
[1]   Visual screening for localized RNAs in yeast revealed novel RNAs at the bud-tip [J].
Andoh, Tomoko ;
Oshiro, Yukiko ;
Hayashi, Sachiko ;
Takeo, Hideki ;
Tani, Tokio .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2006, 351 (04) :999-1004
[2]  
[Anonymous], 1994, METHODS YEAST GENETI
[3]  
[Anonymous], 1989, Molecular Cloning
[4]  
Bailey TL., 1994, P 2 INT C INT SYST M, V2, P28
[5]   A SIMPLE AND EFFICIENT METHOD FOR DIRECT GENE DELETION IN SACCHAROMYCES-CEREVISIAE [J].
BAUDIN, A ;
OZIERKALOGEROPOULOS, O ;
DENOUEL, A ;
LACROUTE, F ;
CULLIN, C .
NUCLEIC ACIDS RESEARCH, 1993, 21 (14) :3329-3330
[6]   The splicing factor BBP interacts specifically with the pre-mRNA branchpoint sequence UACUAAC [J].
Berglund, JA ;
Chua, K ;
Abovich, N ;
Reed, R ;
Rosbash, M .
CELL, 1997, 89 (05) :781-787
[7]   Localization of ASH1 mRNA particles in living yeast [J].
Bertrand, E ;
Chartrand, P ;
Schaefer, M ;
Shenoy, SM ;
Singer, RH ;
Long, RM .
MOLECULAR CELL, 1998, 2 (04) :437-445
[8]   Asymmetric accumulation of ASH1p in postanaphase nuclei depends on a myosin and restricts yeast mating-type switching to mother cells [J].
Bobola, N ;
Jansen, RP ;
Shin, TH ;
Nasmyth, K .
CELL, 1996, 84 (05) :699-709
[9]   She2p, a novel RNA-binding protein tethers ASH1 mRNA to the Myo4p myosin motor via She3p [J].
Böhl, F ;
Kruse, C ;
Frank, A ;
Ferring, D ;
Jansen, RP .
EMBO JOURNAL, 2000, 19 (20) :5514-5524
[10]   HnRNP K: One protein multiple processes [J].
Bomsztyk, K ;
Denisenko, O ;
Ostrowski, J .
BIOESSAYS, 2004, 26 (06) :629-638