Identification of novel factors involved in or regulating initiation of DNA replication by a genome-wide phenotypic screen in Saccharomyces cerevisiae

Lijuan Ma, Yuanliang Zhai, Daorong Feng, Tsz Choi Chan, Yongjun Lu, Xinrong Fu, Jiafeng Wang, Yanhong Chen, Jianna Li, Ke Xu, Chun Liang

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

DNA replication in eukaryotic cells is tightly regulated to ensure faithful inheritance of the genetic material. While the replicators, replication origins and many replication-initiation proteins in Saccharomyces cerevisiae have been identified and extensively studied, the detailed mechanism that controls the initiation of DNA replication is still not well understood. It is likely that some factors involved in or regulating the initiation of DNA replication have not been discovered. To identify novel DNA replication-initiation proteins and their regulators, we developed a sensitive and comprehensive phenotypic screen by combining several established genetic strategies including plasmid loss assays with plasmids containing a single versus multiple replication origins and colony color sectoring assays. We isolated dozen of mutants in previously known initiation proteins and identified several novel factors, including Ctf1p Ctf3p, Ctf4p, Ctf18p, Adk1p and Cdc60p, whose mutants lose plasmid containing a single replication origin at high rates but lose plasmid carrying multiple replication origins at lower rates. We also show that overexpression of replication initiation proteins causes synthetic dosage lethality or growth defects in ctf1 and ctf18 mutants and that Ctf1p and Ctf18p physically interact with ORC, Cdt1p and MCM proteins. Furthermore, depletion of both Ctf1p and Ctf18p prevents S phase entry, retards S phase progression, and reduces pre-RC formation during the M-to-G1 transition. These data suggest that Ctf1p and Ctf18p together play important roles in regulating the initiation of DNA replication.

Original languageEnglish (US)
Pages (from-to)4399-4410
Number of pages12
JournalCell Cycle
Volume9
Issue number21
DOIs
StatePublished - Nov 1 2010
Externally publishedYes

Fingerprint

Replication Origin
DNA Replication
Saccharomyces cerevisiae
Genome
Plasmids
S Phase
Proteins
Saccharomyces cerevisiae Proteins
Eukaryotic Cells
Color
Growth
Genes

Keywords

  • Budding yeast
  • CTF1
  • CTF18
  • DNA replication initiation
  • Phenotypic screen
  • Plasmid loss

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Developmental Biology

Cite this

Identification of novel factors involved in or regulating initiation of DNA replication by a genome-wide phenotypic screen in Saccharomyces cerevisiae. / Ma, Lijuan; Zhai, Yuanliang; Feng, Daorong; Chan, Tsz Choi; Lu, Yongjun; Fu, Xinrong; Wang, Jiafeng; Chen, Yanhong; Li, Jianna; Xu, Ke; Liang, Chun.

In: Cell Cycle, Vol. 9, No. 21, 01.11.2010, p. 4399-4410.

Research output: Contribution to journalArticle

Ma, Lijuan ; Zhai, Yuanliang ; Feng, Daorong ; Chan, Tsz Choi ; Lu, Yongjun ; Fu, Xinrong ; Wang, Jiafeng ; Chen, Yanhong ; Li, Jianna ; Xu, Ke ; Liang, Chun. / Identification of novel factors involved in or regulating initiation of DNA replication by a genome-wide phenotypic screen in Saccharomyces cerevisiae. In: Cell Cycle. 2010 ; Vol. 9, No. 21. pp. 4399-4410.
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AU - Lu, Yongjun

AU - Fu, Xinrong

AU - Wang, Jiafeng

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