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Amidst multiple binding orientations on fork DNA, Saccharolobus MCM helicase proceeds N-first for unwinding.

ABSTRACT: DNA replication requires that the duplex genomic DNA strands be separated; a function that is implemented by ring-shaped hexameric helicases in all Domains. Helicases are composed of two domains, an N- terminal DNA binding domain (NTD) and a C- terminal motor domain (CTD). Replication is controlled by loading of helicases at origins of replication, activation to preferentially encircle one strand, and then translocation to begin separation of the two strands. Using a combination of site-specific DNA footprinting, single-turnover unwinding assays, and unique fluorescence translocation monitoring, we have been able to quantify the binding distribution and the translocation orientation of Saccharolobus (formally Sulfolobus) solfataricus MCM on DNA. Our results show that both the DNA substrate and the C-terminal winged-helix (WH) domain influence the orientation but that translocation on DNA proceeds N-first.

SUBMITTER: Perera HM

PROVIDER: S-EPMC6831031 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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Amidst multiple binding orientations on fork DNA, <i>Saccharolobus</i> MCM helicase proceeds N-first for unwinding.

Perera Himasha M HM Trakselis Michael A MA

eLife 20191029

DNA replication requires that the duplex genomic DNA strands be separated; a function that is implemented by ring-shaped hexameric helicases in all Domains. Helicases are composed of two domains, an N- terminal DNA binding domain (NTD) and a C- terminal motor domain (CTD). Replication is controlled by loading of helicases at origins of replication, activation to preferentially encircle one strand, and then translocation to begin separation of the two strands. Using a combination of site-specific ...[more]

PMID: 31661075

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