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Nucleic Acids Research, 1994, Vol. 22, No. 25 5576-5581
© 1994

Articles

Molecular cloning of MSSP-2, a c-rnyc gene single-strand binding protein: characterization of binding specificity and DNA replication activity

Toshiki Takai, Yoshinori Nishita, Sanae M.M. Iguchi-Ariga1 and Hiroyoshi Ariga*,

Faculty of Pharmaceutical Sciences, Hokkaido University Kita-Ku, Sapporo 060, Japan 1College of Medical Technology, Hokkaido University Kita-Ku, Sapporo 060, Japan

*To whom correspondence should be addressed

Received September 28, 1994. Revised November 18, 1994. Accepted November 18, 1994.

We have previously reported the human cDNA encoding MSSP-1, a sequence-specific double- and single-stranded DNA binding protein [Negishl, Nishita, Sa6gusa, Kaklzakl, Galll, Klhara, Tamal, Miyajlma, Iguchl-Arlga and Arlga (1994) Oncogene, 9, 1133-1143]. MSSP-1 binds to a DNA replication orlgln/transcrlptlonal enhancer of the human c-myc gene and has turned out to be identical with Scr2, a human protein which complements the defect of cdc2 klnase in S.pombe [Kataoka and Nojlma (1994) Nucleic Acid Res., 22, 2687 - 2693]. We have cloned the cDNA for MSSP-2, another member of the MSSP family of proteins. The MSSP-2 cDNA shares highly homologous sequences with MSSP-1 cDNA, except for the Insertion of 48 bp coding 16 amino acids near the C-termlnus. Like MSSP-1, MSSP-2 has RNP-1 consensus sequences. The results of the experiments using bacterlally expressed MSSP-2, and Its deletion mutants, as histldlne fusion proteins suggested that the binding specificity of MSSP-2 to double- and single-stranded DNA is the same as that of MSSP-1, and that the RNP consensus sequences are required for the DNA binding of the protein. MSSP-2 stimulated the DNA replication of an SV40-derived plasmid containing the binding sequence for MSSP-1 or -2. MSSP-2 Is hence suggested to play an important role In regulation of DNA replication.


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