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J. Biol. Chem., Vol. 278, Issue 37, 34983-34989, September 12, 2003

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Sequential Binding of SeqA to Paired Hemi-methylated GATC Sequences Mediates Formation of Higher Order Complexes^*

Joo Seok Han  , Sukhyun Kang  , Ho Lee, Hak Kyun Kim  and Deog Su Hwang ¶

From the Institute of Molecular Biology and Genetics, School of Biological Sciences, Seoul National University, Seoul 151-742, Korea

Preferential binding of the SeqA protein to hemi-methylated GATC sequences functions as a negative regulator for Escherichia coli initiation of chromosomal replication at oriC and is implicated in segregating replicated chromosomes for cell division. We demonstrate that sequential binding of one SeqA tetramer to a set of two hemi-methylated sites mediates formation of higher-order complexes. The absence of cross-binding to separate DNAs suggests that two monomers of a SeqA tetramer bind to two hemi-methylated sites on DNA. The interaction among SeqA proteins bound to at least six adjacent hemi-methylated sites induces aggregation of free proteins to bound proteins. Aggregation might be indicative of SeqA foci, which appear to track replication forks in vivo. Studies of the properties of SeqA binding will contribute to our understanding of the function of SeqA.

Received for publication, May 10, 2003 , and in revised form, June 9, 2003.

^* This work was supported by Grant M102KK010001-02K1101-00820 from the 21C Frontier Microbial Genomics and Applications Center Program, Ministry of Science and Technology and by Grant R02-2002-000-00127-0 and R01-1999-000-00115-0 from the Basic Research Program of the Korea Science and Engineering Foundation and Republic of Korea. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Both authors contributed equally to this work.

Supported by BK21 Research Fellowship from the Ministry of Education, Republic of Korea.

^¶ To whom correspondence should be addressed. E-mail: dshwang{at}plaza.snu.ac.kr.

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