Overexpression of mouse p140 subunit of replication factor C accelerates cellular proliferation.
Screening of a murine cDNA expression library with an IFN-stimulated response element (ISRE), as a recognition site DNA probe, resulted in the isolation of a cDNA encoding a polypeptide of 1145 amino acids designated ISRE-binding factor-1. This was subsequently shown to be identical to the M(r) 140,000 subunit of replication factor C (RFC). RFC is required, along with the proliferating cell nuclear antigen and DNA polymerase delta, for the synthesis of the leading strand during DNA replication. RFC exhibits a structure-specific DNA-binding activity that has been localized to its M(r) 140,000 subunit (p140). Sequence-specific binding of this polypeptide to the ISRE occurs only with low affinity. Based on DNA-binding activity of the truncated RFC-p140 encoded by the partial cDNA isolated, the DNA-binding domain of this polypeptide was mapped to a region encoded by amino acids 366 to 540. Transfection of NIH 3T3 cells with an expression plasmid containing murine RFC-p140 driven by cytomegalovirus early promoter led to the establishment of stable cell lines that expressed a 2.5- to 3.0-fold higher RFC-p140 protein level in comparison with control cells. The stable clones exhibited significantly accelerated cell proliferation, indicating that RFC-p140 is the limiting subunit of an active RFC complex in normal cells.
Jaharul Haque
S
S
Department of Cancer Biology, Cleveland Clinic Foundation, Ohio 44195, USA.
van der Kuip
H
H
Kumar
A
A
Aulitzky
W E
WE
Rutherford
M N
MN
Huber
C
C
Fischer
T
T
Williams
B R
BR
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002453:Cell Cycle; D002455:Cell Division; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D015342:DNA Probes; D004261:DNA Replication; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins; D007372:Interferons / Q000502:physiology; D051379:Mice; D015778:Minor Histocompatibility Antigens; D008969:Molecular Sequence Data; D008970:Molecular Weight; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D051818:Replication Protein C; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014162:Transfection
C096816:BCL2-related protein A1; D015342:DNA Probes; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; C096063:MATA1 protein, S cerevisiae; D015778:Minor Histocompatibility Antigens; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D004247:DNA; D007372:Interferons; D051818:Replication Protein C