2024-03-28T15:04:21Zhttps://eprints.lib.hokudai.ac.jp/dspace-oai/requestoai:eprints.lib.hokudai.ac.jp:2115/540412023-01-19T03:10:04Zhdl_2115_20044hdl_2115_124Molecular cloning of MSSP-2, a c-myc gene single-strand binding protein: characterization of binding specificity and DNA replication activity.Takai, ToshikiNishita, YoshinoriIguchi-Ariga, Sanae M. M.Ariga, Hiroyoshi499Base SequenceCloning, MolecularConsensus SequenceDNA Primers/chemistryDNA ReplicationDNA, Complementary/geneticsDNA-Binding Proteins/geneticsDNA-Binding Proteins/metabolismGenes, mycHumansMolecular Sequence DataMutagenesis, Site-DirectedRNA-Binding ProteinsSequence AlignmentSequence Homology, Amino AcidStructure-Activity RelationshipWe have previously reported the human cDNA encoding MSSP-1, a sequence-specific double- and single-stranded DNA binding protein [Negishi, Nishita, Saƫgusa, Kakizaki, Galli, Kihara, Tamai, Miyajima, Iguchi-Ariga and Ariga (1994) Oncogene, 9, 1133-1143]. MSSP-1 binds to a DNA replication origin/transcriptional 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 kinase in S.pombe [Kataoka and Nojima (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-terminus. Like MSSP-1, MSSP-2 has RNP-1 consensus sequences. The results of the experiments using bacterially expressed MSSP-2, and its deletion mutants, as histidine 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.Oxford University PressJournal Articleapplication/pdfhttp://hdl.handle.net/2115/54041https://eprints.lib.hokudai.ac.jp/dspace/bitstream/2115/54041/1/68_Ariga_1994_Nucleic_Acids_Res.pdf0305-1048Nucleic acids research2225557655811994-12-25enginfo:pmid/7838710info:doi/10.1093/nar/22.25.5576publisher