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The human GINS complex binds to and specifically stimulates human DNA polymerase α‐primase
Author(s) -
De Falco Mariarosaria,
Ferrari Elena,
De Felice Mariarita,
Rossi Mosè,
Hübscher Ulrich,
Pisani Francesca M
Publication year - 2007
Publication title -
embo reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.584
H-Index - 184
eISSN - 1469-3178
pISSN - 1469-221X
DOI - 10.1038/sj.embor.7400870
Subject(s) - primase , dna polymerase , polymerase , replisome , dna clamp , biology , dna polymerase ii , dna , genetics , microbiology and biotechnology , polymerase chain reaction , gene , circular bacterial chromosome , reverse transcriptase
The eukaryotic GINS complex has an essential role in the initiation and elongation phases of genome duplication. It is composed of four paralogous subunits—Sld5, Psf1, Psf2 and Psf3—which are ubiquitous and evolutionarily conserved in eukaryotic organisms. Here, we report the biochemical characterization of the human GINS complex (hGINS). The four hGINS subunits were coexpressed in Escherichia coli in a highly soluble form and purified as a complex. hGINS was shown to interact directly with the heterodimeric human DNA primase, by using either surface plasmon resonance measurements or by immunoprecipitation experiments carried out with anti‐hGINS antibodies. The DNA polymerase α‐primase synthetic activity was specifically stimulated by hGINS on various primed DNA templates. The significance of these findings is discussed in view of the molecular dynamics at the human replication fork.