Functions of sensor 1 and sensor 2 regions of Saccharomyces cerevisiae Cdc6p in vivo and in vitro

Naoko Takahashi, Shinji Tsutsumi, Tomofusa Tsuchiya, Bruce Stillman, Tohru Mizushima

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Cdc6p is a key regulator of the cell cycle in eukaryotes and is a member of the AAA + (ATPases associated with a variety of cellular activities) family of proteins. In this family of proteins, the sensor 1 and sensor 2 regions are important for their function and ATPase activity. Here, site-directed mutagenesis has been used to examine the role of these regions of Saccharomyces cerevisiae Cdc6p in controlling the cell cycle progression and initiation of DNA replication. Two important amino acid residues (Asn 263 in sensor 1 and Arg 332 in sensor 2) were identified as key residues for Cdc6p function in vivo. Cells expressing mutant Cdc6p (N263A or R332E) grew slowly and accumulated in the S phase. In cells expressing mutant Cdc6p, loading of the minichromosome maintenance (MCM) complex of proteins was decreased, suggesting that the slow progression of S phase in these cells was due to inefficient MCM loading on chromatin. Purified wild type Cdc6p but not mutant Cdc6p (N263A and R332E) caused the structural modification of origin recognition complex proteins. These results are consistent with the idea that Cdc6p uses its ATPase activity to change the conformation of origin recognition complex, and then together they recruit the MCM complex.

Original languageEnglish
Pages (from-to)16033-16040
Number of pages8
JournalJournal of Biological Chemistry
Volume277
Issue number18
DOIs
Publication statusPublished - 2002 May 3
Externally publishedYes

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Origin Recognition Complex
Yeast
Saccharomyces cerevisiae
Adenosine Triphosphatases
Cells
S Phase
Cell Cycle
Sensors
Minichromosome Maintenance Proteins
Maintenance
Site-Directed Mutagenesis
Eukaryota
DNA Replication
Mutagenesis
Proteins
Chromatin
Conformations
Amino Acids
In Vitro Techniques
DNA

ASJC Scopus subject areas

  • Biochemistry

Cite this

Functions of sensor 1 and sensor 2 regions of Saccharomyces cerevisiae Cdc6p in vivo and in vitro. / Takahashi, Naoko; Tsutsumi, Shinji; Tsuchiya, Tomofusa; Stillman, Bruce; Mizushima, Tohru.

In: Journal of Biological Chemistry, Vol. 277, No. 18, 03.05.2002, p. 16033-16040.

Research output: Contribution to journalArticle

Takahashi, N, Tsutsumi, S, Tsuchiya, T, Stillman, B & Mizushima, T 2002, 'Functions of sensor 1 and sensor 2 regions of Saccharomyces cerevisiae Cdc6p in vivo and in vitro', Journal of Biological Chemistry, vol. 277, no. 18, pp. 16033-16040. https://doi.org/10.1074/jbc.M108615200
Takahashi, Naoko ; Tsutsumi, Shinji ; Tsuchiya, Tomofusa ; Stillman, Bruce ; Mizushima, Tohru. / Functions of sensor 1 and sensor 2 regions of Saccharomyces cerevisiae Cdc6p in vivo and in vitro. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 18. pp. 16033-16040.
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