PSF1 Is Essential for Early Embryogenesis in Mice

ABSTRACT Psf1 (partner of sld five 1) forms a novel heterotetramer complex, GINS (Go, Ichi, Nii, and San; five, one, two, and three, respectively, in Japanese), with Sld5, Psf2, and Psf3. The formation of this complex is essential for the initiation of DNA replication in yeast and Xenopus laevis egg extracts. Although all of the components are well conserved in higher eukaryotes, the biological function in vivo is largely unknown. We originally cloned the mouse ortholog of PSF1 from a hematopoietic stem cell cDNA library and found that PSF1 is expressed in blastocysts, adult bone marrow, and testis, in which the stem cell system is active. Here we used the gene-targeting technique to determine the physiological function of PSF1 in vivo. Mice homozygous for a nonfunctional mutant of PSF1 died in utero around the time of implantation. PSF1− / − blastocysts failed to show outgrowth in culture and exhibited a cell proliferation defect. Our data clearly indicate that PSF1 is required for early embryogenesis.

[1]  M. Méchali,et al.  Vertebrate HoxB gene expression requires DNA replication , 2003, The EMBO journal.

[2]  S. Forlani,et al.  Relief of a repressed gene expression state in the mouse 1-cell embryo requires DNA replication. , 1998, Development.

[3]  Ronald W. Davis,et al.  Functional characterization of the S. cerevisiae genome by gene deletion and parallel analysis. , 1999, Science.

[4]  B. Stillman,et al.  The Cdc6p nucleotide-binding motif is required for loading mcm proteins onto chromatin. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Anindya Dutta,et al.  Requirement of CDC45 for Postimplantation Mouse Development , 2001, Molecular and Cellular Biology.

[6]  B. Stillman,et al.  Human Orc2 localizes to centrosomes, centromeres and heterochromatin during chromosome inheritance , 2005, The EMBO journal.

[7]  J. Diffley,et al.  Two steps in the assembly of complexes at yeast replication origins in vivo , 1994, Cell.

[8]  J. Roder,et al.  Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[9]  H. Takisawa,et al.  Xenopus Cdc45‐dependent loading of DNA polymerase α onto chromatin under the control of S‐phase cdk , 1998, The EMBO journal.

[10]  Anindya Dutta,et al.  DNA replication in eukaryotic cells. , 2002, Annual review of biochemistry.

[11]  R. Schultz.  Regulation of zygotic gene activation in the mouse , 1993, BioEssays : news and reviews in molecular, cellular and developmental biology.

[12]  T. Kelly,et al.  Regulation of chromosome replication. , 2000, Annual review of biochemistry.

[13]  J A Wohlschlegel,et al.  Inhibition of eukaryotic DNA replication by geminin binding to Cdt1. , 2000, Science.

[14]  Hiroyuki Araki,et al.  GINS, a novel multiprotein complex required for chromosomal DNA replication in budding yeast. , 2003, Genes & development.

[15]  Ronald W. Davis,et al.  Replication dynamics of the yeast genome. , 2001, Science.

[16]  Masato T. Kanemaki,et al.  Functional proteomic identification of DNA replication proteins by induced proteolysis in vivo , 2003, Nature.

[17]  Charles Kooperberg,et al.  Genome-wide DNA replication profile for Drosophila melanogaster: a link between transcription and replication timing , 2002, Nature Genetics.

[18]  H. Araki,et al.  A novel ring-like complex of Xenopus proteins essential for the initiation of DNA replication. , 2003, Genes & development.

[19]  J. Diffley,et al.  DNA synthesis at individual replication forks requires the essential initiation factor Cdc45p , 2000, The EMBO journal.

[20]  O. Aparicio,et al.  Components and Dynamics of DNA Replication Complexes in S. cerevisiae: Redistribution of MCM Proteins and Cdc45p during S Phase , 1997, Cell.

[21]  B. Tye,et al.  Drosophila MCM10 interacts with members of the prereplication complex and is required for proper chromosome condensation. , 2003, Molecular biology of the cell.

[22]  S. Dalton,et al.  Cdc45p assembles into a complex with Cdc46p/Mcm5p, is required for minichromosome maintenance, and is essential for chromosomal DNA replication. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[23]  G. Yancopoulos,et al.  Critical role of the TIE2 endothelial cell receptor in the development of definitive hematopoiesis. , 1998, Immunity.

[24]  T. Noda,et al.  A Role for Hematopoietic Stem Cells in Promoting Angiogenesis , 2000, Cell.