Pir1p mediates translocation of the yeast Apn1p endonuclease into the mitochondria to maintain genomic stability

被引:84
作者
Vongsamphanh, R [1 ]
Fortier, PK [1 ]
Ramotar, D [1 ]
机构
[1] Univ Montreal, Guy Bernier Res Ctr, Montreal, PQ H1T 2M4, Canada
关键词
D O I
10.1128/MCB.21.5.1647-1655.2001
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 mitochondrial genome is continuously subject to attack by reactive oxygen species generated through aerobic metabolism. This leads to the formation of a variety of highly genotoxic DNA lesions, including abasic sites. Yeast Apn1p is localized to the nucleus, where it functions to cleave abasic sites, and apn1 Delta mutants are hypersensitive to agents such as methyl methanesulfonate (MR;IS) that induce abasic sites. Here we demonstrate for the first time that yeast Apn1p is also localized to the mitochondria. We found that Pir1p, initially isolated as a cell wall constituent of unknown function, interacts with the C-terminal end of Apn1p, which bears a bipartite nuclear localization signal. Further analysis revealed that Pir1p is required to cause Apn1p mitochondrial localization, presumably by competing with the nuclear transport machinery, pir1 Delta mutants displayed a striking (similar to3-fold) increase of Apn1p in the nucleus, which coincided with drastically reduced levels in the mitochondria. To explore the functional consequences of the Apn1p-Pir1p interaction, we measured the rate of mitochondrial mutations in the wild type and pir1 Delta and apn1 Delta mutants. pir1 Delta and apn1 Delta mutants exposed to MMS exhibited 3.6- and 5.8-fold increases, respectively, in the rate of mitochondrial mutations, underscoring the importance of Apn1p in repair of the mitochondrial genome. We conclude that Pir1p interacts with Apn1p, at the level of either the cytoplasm or nucleus, and facilitates Apn1p transport into the mitochondria to repair damaged DNA.
引用
收藏
页码:1647 / 1655
页数:9
相关论文
共 52 条
[1]   Mitochondrial DNA polymorphisms in pathologically proven Parkinson's disease [J].
Bandmann, O ;
Sweeney, MG ;
Daniel, SE ;
Marsden, CD ;
Wood, NW .
JOURNAL OF NEUROLOGY, 1997, 244 (04) :262-265
[2]   Crystallographic analysis of the recognition of a nuclear localization signal by the nuclear import factor karyopherin α [J].
Conti, E ;
Uy, M ;
Leighton, L ;
Blobel, G ;
Kuriyan, J .
CELL, 1998, 94 (02) :193-204
[3]   Overexpression of yeast karyopherin Pse1p/Kap121p stimulates the mitochondrial import of hydrophobic proteins in vivo [J].
Corral-Debrinski, M ;
Belgareh, N ;
Blugeon, C ;
Claros, MG ;
Doye, V ;
Jacq, C .
MOLECULAR MICROBIOLOGY, 1999, 31 (05) :1499-1511
[4]   An oxidative damage-specific endonuclease from rat liver mitochondria [J].
Croteau, DL ;
apRhys, CMJ ;
Hudson, EK ;
Dianov, GL ;
Hansford, RG ;
Bohr, VA .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (43) :27338-27344
[5]   A SINGLE NUCLEOTIDE SUBSTITUTION AT THE RIB2 LOCUS OF THE YEAST MITOCHONDRIAL GENE FOR 21S RIBOSOMAL-RNA CONFERS RESISTANCE TO ERYTHROMYCIN AND COLD-SENSITIVE RIBOSOME ASSEMBLY [J].
CUI, Z ;
MASON, TL .
CURRENT GENETICS, 1989, 16 (04) :273-279
[6]   RETRACTED: Mutations in mitochondrial cytochrome c oxidase genes segregate with late-onset Alzheimer disease (Retracted Article) [J].
Davis, RE ;
Miller, S ;
Herrnstadt, C ;
Ghosh, SS ;
Fahy, E ;
Shinobu, LA ;
Galasko, D ;
Thal, LJ ;
Beal, MF ;
Howell, N ;
Parker, WD .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (09) :4526-4531
[7]  
Dove JE, 1998, METHOD CELL BIOL, V53, P33
[8]   PROTEIN BLOTTING - PRINCIPLES AND APPLICATIONS [J].
GERSHONI, JM ;
PALADE, GE .
ANALYTICAL BIOCHEMISTRY, 1983, 131 (01) :1-15
[9]   NOVEL MUTAGENIC PROPERTIES OF ABASIC SITES IN SACCHAROMYCES-CEREVISIAE [J].
GIBBS, PEM ;
LAWRENCE, CW .
JOURNAL OF MOLECULAR BIOLOGY, 1995, 251 (02) :229-236
[10]   APPLICATIONS OF HIGH-EFFICIENCY LITHIUM-ACETATE TRANSFORMATION OF INTACT YEAST-CELLS USING SINGLE-STRANDED NUCLEIC-ACIDS AS CARRIER [J].
GIETZ, RD ;
SCHIESTL, RH .
YEAST, 1991, 7 (03) :253-263