Mitochondrial DNA ligase function in Saccharomyces cerevisiae

被引:24
作者
Donahue, SL
Corner, BE
Bordone, L
Campbell, C
机构
[1] Univ Minnesota, Sch Med, Dept Pharmacol, Minneapolis, MN 55455 USA
[2] Univ Minnesota, Dept Plant Biol, St Paul, MN 55108 USA
关键词
D O I
10.1093/nar/29.7.1582
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 Saccharomyces cerevisiae CDC9 gene encodes a DNA ligase protein that is targeted to both the nucleus and the mitochondria, While nuclear Cdc9p is known to play an essential role in nuclear DNA replication and repair, its role in mitochondrial DNA dynamics has not been defined, It is also unclear whether additional DNA ligase proteins are present in yeast mitochondria, To address these issues, mitochondrial DNA ligase function in S.cerevisiae was analyzed. Biochemical analysis of mitochondrial protein extracts supported the conclusion that Cdc9p was the sole DNA ligase protein present in this organelle. Inactivation of mitochondrial Cdc9p function led to a rapid decline in cellular mitochondrial DNA content in both dividing and stationary yeast cultures. In contrast, there was no apparent defect in mitochondrial DNA dynamics in a yeast strain deficient in Dnl4p (Delta dnl4). The Escherichia coli EcoRI endonuclease was targeted to yeast mitochondria, Transient expression of this recombinant EcoRI endonuclease led to the formation of mitochondrial DNA double-strand breaks, While wild-type and Delta dnl4 yeast were able to rapidly recover from this mitochondrial DNA damage, clones deficient in mitochondrial Cdc9p were not. These results support the conclusion that yeast rely upon a single DNA ligase, Cdc9p, to carry out mitochondrial DNA replication and recovery from both spontaneous and induced mitochondrial DNA damage.
引用
收藏
页码:1582 / 1589
页数:8
相关论文
共 35 条
[1]  
Adams A., 1997, METHODS YEAST GENETI
[2]  
[Anonymous], METHOD ENZYMOL
[3]  
ARRAND JE, 1986, J BIOL CHEM, V261, P9079
[4]   AN IMPROVED ASSAY FOR DNA-LIGASE REVEALS TEMPERATURE-SENSITIVE ACTIVITY IN CDC9 MUTANTS OF SACCHAROMYCES-CEREVISIAE [J].
BARKER, DG ;
JOHNSON, AL ;
JOHNSTON, LH .
MOLECULAR & GENERAL GENETICS, 1985, 200 (03) :458-462
[5]   REGULATED EXPRESSION OF ENDONUCLEASE ECORI IN SACCHAROMYCES-CEREVISIAE - NUCLEAR ENTRY AND BIOLOGICAL CONSEQUENCES [J].
BARNES, G ;
RINE, J .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1985, 82 (05) :1354-1358
[6]   DNA double-strand-break sensitivity, DNA replication, and cell cycle arrest phenotypes of Ku-deficient Saccharomyces cerevisiae [J].
Barnes, G ;
Rio, D .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (03) :867-872
[7]  
Butow R A, 1996, Methods Enzymol, V264, P265, DOI 10.1016/S0076-6879(96)64026-9
[8]  
CALECOTT KW, 1996, NUCLEIC ACIDS RES, V24, P4387
[9]   FACS-optimized mutants of the green fluorescent protein (GFP) [J].
Cormack, BP ;
Valdivia, RH ;
Falkow, S .
GENE, 1996, 173 (01) :33-38
[10]  
Dujon B., 1981, MOL BIOL YEAST SACCH, P505