The yeast Dbf4 Zn2+ finger domain suppresses single-stranded DNA at replication forks initiated from a subset of origins

Dbf4 is the cyclin-like subunit for the Dbf4-dependent protein kinase (DDK), required for activating the replicative helicase at DNA replication origin that fire during S phase. Dbf4 also functions as an adaptor, targeting the DDK to different groups of origins and substrates. Here we report a genom...

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Bibliographic Details
Published in:Current genetics 2022-04, Vol.68 (2), p.253-265
Main Authors: Bachant, Jeff, Hoffman, Elizabeth A., Caridi, Chris, Nugent, Constance I., Feng, Wenyi
Format: Article
Language:English
Subjects:
Accumulation
Biochemistry
Biomedical and Life Sciences
C-Terminus
Cell Biology
Deoxyribonucleic acid
DNA
DNA biosynthesis
DNA helicase
DNA primase
Domains
Emission
Genomes
Kinases
Life Sciences
Microbial Genetics and Genomics
Microbiology
Mutants
Original
Original Article
Origins
Phenotypes
Plant Sciences
Primase
Protein kinase
Proteomics
Replication
Replication forks
Replication origins
Restarting
S phase
Single-stranded DNA
Substrates
Yeast
Yeasts
Zinc
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Summary:Dbf4 is the cyclin-like subunit for the Dbf4-dependent protein kinase (DDK), required for activating the replicative helicase at DNA replication origin that fire during S phase. Dbf4 also functions as an adaptor, targeting the DDK to different groups of origins and substrates. Here we report a genome-wide analysis of origin firing in a budding yeast mutant, dbf4-zn , lacking the Zn 2+ finger domain within the C-terminus of Dbf4. At one group of origins, which we call dromedaries, we observe an unanticipated DNA replication phenotype: accumulation of single-stranded DNA spanning ± 5kbp from the center of the origins. A similar accumulation of single-stranded DNA at origins occurs more globally in pri1-m4 mutants defective for the catalytic subunit of DNA primase and rad53 mutants defective for the S phase checkpoint following DNA replication stress. We propose the Dbf4 Zn 2+ finger suppresses single-stranded gaps at replication forks emanating from dromedary origins. Certain origins may impose an elevated requirement for the DDK to fully initiate DNA synthesis following origin activation. Alternatively, dbf4-zn may be defective for stabilizing/restarting replication forks emanating from dromedary origins during replication stress.
ISSN:0172-8083
1432-0983
DOI:10.1007/s00294-022-01230-6