Functional analysis of CedA based on its structure: Residues important in binding of DNA and RNA polymerase and in the cell division regulation

Yoshito Abe, Naoki Fujisaki, Takanori Miyoshi, Noriko Watanabe, Tsutomu Katayama, Tadashi Ueda

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

DnaAcos, a mutant of the initiator DnaA, causes overinitiation of chromosome replication in Escherichia coli, resulting in inhibition of cell division. CedA was found to be a multi-copy suppressor which represses the dnaAcos inhibition of cell division. However, functional mechanism of CedA remains elusive except for previously indicated possibilities in binding to DNA and RNA polymerase. In this study, we searched for the specific sites of CedA in binding of DNA and RNA polymerase and in repression of cell division inhibition. First, DNA sequence to which CedA preferentially binds was determined. Next, the several residues and β4 region in CedA C-terminal domain was suggested to specifically interact with the DNA. Moreover, we found that the flexible N-terminal region was required for tight binding to longer DNA as well as interaction with RNA polymerase. Based on these results, several cedA mutants were examined in ability for repressing dnaAcos cell division inhibition. We found that the N-terminal region was dispensable and that Glu32 in the C-terminal domain was required for the repression. These results suggest that CedA has multiple roles and residues with different functions are positioned in the two regions.

Original languageEnglish
Pages (from-to)217-223
Number of pages7
JournalJournal of biochemistry
Volume159
Issue number2
DOIs
Publication statusPublished - May 13 2015

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Functional analysis
DNA-Directed DNA Polymerase
DNA-Directed RNA Polymerases
Cell Division
Cells
DNA sequences
DNA
Chromosomes
Escherichia coli

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Functional analysis of CedA based on its structure : Residues important in binding of DNA and RNA polymerase and in the cell division regulation. / Abe, Yoshito; Fujisaki, Naoki; Miyoshi, Takanori; Watanabe, Noriko; Katayama, Tsutomu; Ueda, Tadashi.

In: Journal of biochemistry, Vol. 159, No. 2, 13.05.2015, p. 217-223.

Research output: Contribution to journalArticle

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