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this report, in order to clarify the mode of action of the Mcm helicase and obtain insight into how it may function at the replication forks in vivo, we have conducted detailed analyses of helicase action and DNA binding of mouse Mcm4/6/7 helicase using various forked and bubble substrate DNAs. We specifically addressed sequence requirement for the Mcm helicase activation, mode of interaction with DNA substrates and whether the thymine sequences are required for continuous activation of Mcm helicase during the unwinding process. The results indicate that mammalian Mcm4/6/7 primarily binds to single-stranded DNA region, and that the extent of helicase activation is related to the thymine content of the single-stranded segment. Unexpectedly, Mcm4/6/7 helicase is not capable of efficiently unwinding the GC-rich duplex segment, suggesting that some other mechanism may be required for completion of replication of the entire genome."}

{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T722","span":{"begin":62,"end":65},"obj":"GO:0042555"},{"id":"T723","span":{"begin":66,"end":74},"obj":"GO_EXT:0004386"},{"id":"T724","span":{"begin":126,"end":137},"obj":"GO:0006260"},{"id":"T725","span":{"begin":126,"end":143},"obj":"GO:0005657"},{"id":"T726","span":{"begin":192,"end":200},"obj":"GO_EXT:0004386"},{"id":"T727","span":{"begin":212,"end":215},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T728","span":{"begin":216,"end":223},"obj":"CHEMINF_GO_EXT:chemical_binding_or_bond_formation"},{"id":"T729","span":{"begin":227,"end":232},"obj":"NCBITaxon:10088"},{"id":"T730","span":{"begin":233,"end":237},"obj":"PR_EXT:000010246"},{"id":"T731","span":{"begin":233,"end":241},"obj":"GO:0097373"},{"id":"T732","span":{"begin":242,"end":250},"obj":"GO_EXT:0004386"},{"id":"T733","span":{"begin":265,"end":271},"obj":"GO:0005657"},{"id":"T734","span":{"begin":293,"end":297},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T735","span":{"begin":325,"end":333},"obj":"SO_EXT:biological_sequence"},{"id":"T736","span":{"begin":354,"end":357},"obj":"GO:0042555"},{"id":"T737","span":{"begin":358,"end":366},"obj":"GO_EXT:0004386"},{"id":"T738","span":{"begin":404,"end":407},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T739","span":{"begin":435,"end":442},"obj":"CHEBI_EXT:thymine_nucleobase_or_nucleoside_or_nucleotide_molecular_entity_or_group"},{"id":"T740","span":{"begin":443,"end":452},"obj":"SO_EXT:biological_sequence"},{"id":"T741","span":{"begin":495,"end":498},"obj":"GO:0042555"},{"id":"T742","span":{"begin":499,"end":507},"obj":"GO_EXT:0004386"},{"id":"T743","span":{"begin":564,"end":573},"obj":"NCBITaxon:40674"},{"id":"T744","span":{"begin":574,"end":578},"obj":"PR_EXT:000010246"},{"id":"T745","span":{"begin":574,"end":582},"obj":"GO:0097373"},{"id":"T746","span":{"begin":593,"end":598},"obj":"CHEMINF_GO_EXT:chemical_binding_or_bond_formation"},{"id":"T747","span":{"begin":602,"end":621},"obj":"CHEBI_SO_EXT:single_stranded_DNA"},{"id":"T748","span":{"begin":653,"end":661},"obj":"GO_EXT:0004386"},{"id":"T749","span":{"begin":691,"end":698},"obj":"CHEBI_EXT:thymine_nucleobase_or_nucleoside_or_nucleotide_molecular_entity_or_group"},{"id":"T750","span":{"begin":714,"end":729},"obj":"SO:0000984"},{"id":"T751","span":{"begin":753,"end":757},"obj":"PR_EXT:000010246"},{"id":"T752","span":{"begin":753,"end":761},"obj":"GO:0097373"},{"id":"T753","span":{"begin":762,"end":770},"obj":"GO_EXT:0004386"},{"id":"T754","span":{"begin":823,"end":829},"obj":"SO:0000985"},{"id":"T755","span":{"begin":910,"end":921},"obj":"GO:0006260"},{"id":"T756","span":{"begin":936,"end":942},"obj":"SO_EXT:0001026"}],"text":"In this report, in order to clarify the mode of action of the Mcm helicase and obtain insight into how it may function at the replication forks in vivo, we have conducted detailed analyses of helicase action and DNA binding of mouse Mcm4/6/7 helicase using various forked and bubble substrate DNAs. We specifically addressed sequence requirement for the Mcm helicase activation, mode of interaction with DNA substrates and whether the thymine sequences are required for continuous activation of Mcm helicase during the unwinding process. The results indicate that mammalian Mcm4/6/7 primarily binds to single-stranded DNA region, and that the extent of helicase activation is related to the thymine content of the single-stranded segment. Unexpectedly, Mcm4/6/7 helicase is not capable of efficiently unwinding the GC-rich duplex segment, suggesting that some other mechanism may be required for completion of replication of the entire genome."}

{"project":"craft-ca-core-dev","denotations":[{"id":"T604","span":{"begin":62,"end":65},"obj":"GO:0042555"},{"id":"T605","span":{"begin":126,"end":137},"obj":"GO:0006260"},{"id":"T606","span":{"begin":126,"end":143},"obj":"GO:0005657"},{"id":"T607","span":{"begin":227,"end":232},"obj":"NCBITaxon:10088"},{"id":"T608","span":{"begin":233,"end":237},"obj":"PR:000010246"},{"id":"T609","span":{"begin":233,"end":241},"obj":"GO:0097373"},{"id":"T610","span":{"begin":265,"end":271},"obj":"GO:0005657"},{"id":"T611","span":{"begin":354,"end":357},"obj":"GO:0042555"},{"id":"T612","span":{"begin":495,"end":498},"obj":"GO:0042555"},{"id":"T613","span":{"begin":564,"end":573},"obj":"NCBITaxon:40674"},{"id":"T614","span":{"begin":574,"end":578},"obj":"PR:000010246"},{"id":"T615","span":{"begin":574,"end":582},"obj":"GO:0097373"},{"id":"T616","span":{"begin":602,"end":617},"obj":"SO:0000984"},{"id":"T617","span":{"begin":714,"end":729},"obj":"SO:0000984"},{"id":"T618","span":{"begin":753,"end":757},"obj":"PR:000010246"},{"id":"T619","span":{"begin":753,"end":761},"obj":"GO:0097373"},{"id":"T620","span":{"begin":823,"end":829},"obj":"SO:0000985"},{"id":"T621","span":{"begin":910,"end":921},"obj":"GO:0006260"},{"id":"T622","span":{"begin":936,"end":942},"obj":"SO:0001026"}],"text":"In this report, in order to clarify the mode of action of the Mcm helicase and obtain insight into how it may function at the replication forks in vivo, we have conducted detailed analyses of helicase action and DNA binding of mouse Mcm4/6/7 helicase using various forked and bubble substrate DNAs. We specifically addressed sequence requirement for the Mcm helicase activation, mode of interaction with DNA substrates and whether the thymine sequences are required for continuous activation of Mcm helicase during the unwinding process. The results indicate that mammalian Mcm4/6/7 primarily binds to single-stranded DNA region, and that the extent of helicase activation is related to the thymine content of the single-stranded segment. Unexpectedly, Mcm4/6/7 helicase is not capable of efficiently unwinding the GC-rich duplex segment, suggesting that some other mechanism may be required for completion of replication of the entire genome."}