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DNA helicase gene interaction network defined using synthetic lethality analyzed by microarray
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DOI：10.1038/ng1258.png)
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摘要
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We describe a new synthetic lethality analysis by microarray (SLAM) technique that uses 4,600 Saccharomyces cerevisiae haploid deletion mutants with molecular bar codes (TAGs). We used SGS1 and SRS2, two 3' --> 5' DNA helicase genes, as queries to identify their redundant and unique biological functions. We introduced these query mutations into a haploid deletion pool by integrative transformation to disrupt the query gene in every cell, generating a double mutant pool. Optimization of integrative transformation efficiency was essential to the success of SLAM. Synthetic interactions defined a DNA helicase genetic network and predicted a role for SRS2 in processing damaged replication forks but, unlike SGS1, not in rDNA replication, DNA topology or lagging strand synthesis. SGS1 and SRS2 have synthetic defects with MRC1 but not RAD9, suggesting that SGS1 and SRS2 function in a parallel pathway with MRC1 to transduce the DNA replication stress signal to the general DNA damage checkpoint pathway. Both helicase genes have rad51-reversible synthetic defects with 5 3 DNA helicase RRM3, suggesting that RRM3 helps prevent formation of toxic recombination intermediates. SLAM detects synthetic lethality efficiently and ranks candidate genetic interactions, making it an especially useful method.
Keyword:
SACCHAROMYCES-CEREVISIAE GENOME
REPLICATION FORK PROGRESSION
HOMOLOGOUS RECOMBINATION
YEAST SGS1
SRS2
REQUIREMENT
ABSENCE
MUTANTS
PURIFICATION
CHECKPOINT
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IF:
40
论文数:
1.0W
被引数:
10.5W
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