The human GINS complex binds to and specifically stimulates human DNA polymerase alpha-primase

Mariarosaria De Falco; Elena Ferrari; Mariarita De Felice; Mosè Rossi; Ulrich Hübscher; Francesca M Pisani

doi:10.1038/sj.embor.7400870

The human GINS complex binds to and specifically stimulates human DNA polymerase alpha-primase

EMBO Rep. 2007 Jan;8(1):99-103. doi: 10.1038/sj.embor.7400870. Epub 2006 Dec 15.

Authors

Mariarosaria De Falco¹, Elena Ferrari, Mariarita De Felice, Mosè Rossi, Ulrich Hübscher, Francesca M Pisani

Affiliation

¹ Istituto di Biochimica delle Proteine, Consiglio Nazionale delle Ricerche, Via P. Castellino, 111, Napoli 80131, Italy.

Abstract

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 alpha-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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ATP Binding Cassette Transporter, Subfamily B, Member 2
ATP Binding Cassette Transporter, Subfamily B, Member 3
ATP-Binding Cassette Transporters / genetics
ATP-Binding Cassette Transporters / metabolism*
DNA / chemistry
DNA Polymerase I / metabolism*
DNA Primase / metabolism*
DNA Replication
Escherichia coli / genetics
Humans
Peptide Fragments / genetics
Peptide Fragments / metabolism*
Protein Precursors / genetics
Protein Precursors / metabolism*
Protein Subunits / genetics
Protein Subunits / metabolism
Recombinant Proteins / biosynthesis
Recombinant Proteins / genetics
Recombinant Proteins / isolation & purification
Surface Plasmon Resonance

Substances

ATP Binding Cassette Transporter, Subfamily B, Member 2
ATP Binding Cassette Transporter, Subfamily B, Member 3
ATP-Binding Cassette Transporters
Peptide Fragments
Protein Precursors
Protein Subunits
Recombinant Proteins
TAP1 protein, human
thyrotropin-releasing hormone-associated peptide 3, human
TAP2 protein, human
DNA
DNA Primase
DNA polymerase alpha-primase
DNA Polymerase I