TY - JOUR
T1 - Role of the amino-terminal region of the DnaA protein in opening of the duplex DNA at the oriC region
AU - Mima, Shinji
AU - Yamagachi, Yoshihiro
AU - Kondo, Taemi
AU - Tsuchiya, Tomofusa
AU - Mizushima, Tohru
N1 - Funding Information:
We thank Dr T. Ogawa (Nagoya University) and T. Katayama (Kyushu University) for giving us HU protein and the plasmid (M13E10), respectively. We thank also Dr V. Ellison (Cold Spring Harbor Laboratory) for helpful comments on this manuscript. This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture, Japan and the Naito foundation. This study is carried out as a part of ‘Ground Research Announcement for Space Utilization’ promoted by the Japan Space Forum.
PY - 1999/7/1
Y1 - 1999/7/1
N2 - We report in this paper that the amino acid residues Ile-26 and Leu-40 of the DnaA protein are essential for the DNA replication activity in vitro. Lines of evidence to support this conclusion are as follows. Variants of the DnaA protein containing either an Ile-26-Ser or Leu-40-Ser replacement were unable to support oriC DNA replication in vitro. Though the mutant DnaA proteins retained the capability to bind oriC DNA, they were unable to open the duplex DNA at oriC. Based on these and other results, we conclude that the N-terminal region of the DnaA, protein is involved in the oligomerization of this protein, an essential step for the duplex opening activity at oriC.
AB - We report in this paper that the amino acid residues Ile-26 and Leu-40 of the DnaA protein are essential for the DNA replication activity in vitro. Lines of evidence to support this conclusion are as follows. Variants of the DnaA protein containing either an Ile-26-Ser or Leu-40-Ser replacement were unable to support oriC DNA replication in vitro. Though the mutant DnaA proteins retained the capability to bind oriC DNA, they were unable to open the duplex DNA at oriC. Based on these and other results, we conclude that the N-terminal region of the DnaA, protein is involved in the oligomerization of this protein, an essential step for the duplex opening activity at oriC.
KW - DNA replication
KW - DnaA
KW - N-terminal region
KW - Site-directed mutagenesis
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U2 - 10.1016/S0378-1097(99)00233-5
DO - 10.1016/S0378-1097(99)00233-5
M3 - Article
C2 - 10418142
AN - SCOPUS:0032790811
VL - 176
SP - 163
EP - 167
JO - FEMS Microbiology Letters
JF - FEMS Microbiology Letters
SN - 0378-1097
IS - 1
ER -