Role of the rab GTP-binding protein Ypt3 in the fission yeast exocytic pathway and its connection to calcineurin function

被引:81
作者
Cheng, H
Sugiura, R
Wu, WL
Fujita, M
Lu, YB
Sio, SO
Kawai, R
Takegawa, K
Shuntoh, H
Kuno, T [1 ]
机构
[1] Kobe Univ, Grad Sch Med, Div Mol Pharmacol & Pharmacogenom, Dept Genome Sci, Kobe, Hyogo 6500017, Japan
[2] Kagawa Univ, Dept Life Sci, Fac Agr, Kagawa 7610795, Japan
[3] Kobe Univ, Sch Med, Fac Hlth Sci, Kobe, Hyogo 6500142, Japan
关键词
D O I
10.1091/mbc.01-09-0463
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
A genetic screen for mutations synthetically lethal with fission yeast calcineurin deletion led to the identification of Ypt3, a homolog of mammalian Rab11 GTP-binding protein. A mutant with the temperature-sensitive ypt3-i5 allele showed pleiotropic phenotypes such as defects in cytokinesis, cell wall integrity, and vacuole fusion, and these were exacerbated by FK506-treatment, a specific inhibitor of calcineurin. Green fluorescent protein (GFP)-tagged Ypt3 showed cytoplasmic staining that was concentrated at growth sites, and this polarized localization required the actin cytoskeleton. It was also detected as a punctate staining in an actin-independent manner. Electron microscopy revealed that ypt3-i5 mutants accumulated aberrant Golgi-like structures and putative post-Golgi vesicles, which increased remarkably at the restrictive temperature. Consistently, the secretion of GFP fused with the pho1(+) leader peptide (SPL-GFP) was abolished at the restrictive temperature in ypt3-i5 mutants. FK506-treatment accentuated the accumulation of aberrant Golgi-like structures and caused a significant decrease of SPL-GFP secretion at a permissive temperature. These results suggest that Ypt3 is required at multiple steps of the exocytic pathway and its mutation affects diverse cellular processes and that calcineurin is functionally connected to these cellular processes.
引用
收藏
页码:2963 / 2976
页数:14
相关论文
共 58 条
[1]   A NEW TROPOMYOSIN ESSENTIAL FOR CYTOKINESIS IN THE FISSION YEAST S-POMBE [J].
BALASUBRAMANIAN, MK ;
HELFMAN, DM ;
HEMMINGSEN, SM .
NATURE, 1992, 360 (6399) :84-87
[2]  
Balasubramanian MK, 1997, METHOD ENZYMOL, V283, P494
[3]   THE SCHIZOSACCHAROMYCES-POMBE CDC3+ GENE ENCODES A PROFILIN ESSENTIAL FOR CYTOKINESIS [J].
BALASUBRAMANIAN, MK ;
HIRANI, BR ;
BURKE, JD ;
GOULD, KL .
JOURNAL OF CELL BIOLOGY, 1994, 125 (06) :1289-1301
[4]   TATA BOX MUTATIONS IN THE SCHIZOSACCHAROMYCES-POMBE NMT-1 PROMOTER AFFECT TRANSCRIPTION EFFICIENCY BUT NOT THE TRANSCRIPTION START POINT OR THIAMINE REPRESSIBILITY [J].
BASI, G ;
SCHMID, E ;
MAUNDRELL, K .
GENE, 1993, 123 (01) :131-136
[5]   CONSTRUCTION OF A SCHIZOSACCHAROMYCES-POMBE GENE BANK IN A YEAST BACTERIAL SHUTTLE VECTOR AND ITS USE TO ISOLATE GENES BY COMPLEMENTATION [J].
BEACH, D ;
PIPER, M ;
NURSE, P .
MOLECULAR & GENERAL GENETICS, 1982, 187 (02) :326-329
[6]   Two GTPase isoforms, ypt31p and ypt32p, are essential for Golgi function in yeast [J].
Benli, M ;
Doring, F ;
Robinson, DG ;
Yang, XP ;
Gallwitz, D .
EMBO JOURNAL, 1996, 15 (23) :6460-6475
[7]   A selective transport route from golgi to late endosomes that requires the yeast GGA proteins [J].
Black, MW ;
Pelham, HRB .
JOURNAL OF CELL BIOLOGY, 2000, 151 (03) :587-600
[8]   A genomic perspective on membrane compartment organization [J].
Bock, JB ;
Matern, HT ;
Peden, AA ;
Scheller, RH .
NATURE, 2001, 409 (6822) :839-841
[9]   Regulated vacuole fusion and fission in Schizosaccharomyces pombe:: an osmotic response dependent on MAP kinases [J].
Bone, N ;
Millar, JBA ;
Toda, T ;
Armstrong, J .
CURRENT BIOLOGY, 1998, 8 (03) :135-144
[10]   Secretion of heterologous proteins from Schizosaccharomyces pombe using the homologous leader sequence of pho1+ acid phosphatase [J].
Braspenning, J ;
Meschede, W ;
Marchini, A ;
Müller, M ;
Gissmann, L ;
Tommasino, M .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1998, 245 (01) :166-171