TY - JOUR
T1 - HRAD17, a structural homolog of the Schizosaccharomyces pombe RAD17 cell cycle checkpoint gene, stimulates p53 accumulation
AU - Li, Lei
AU - Peterson, Carolyn A.
AU - Kanter-Smoler, Gunilla
AU - Wei, Ying Fei
AU - Ramagli, Louis S.
AU - Sunnerhagen, Per
AU - Siciliano, Michael J.
AU - Legerski, Randy J.
N1 - Funding Information:
This work was supported by grant CA76172 (Lei Li), CA52461 (Randy J Legerski), and CA34936 (Michael J Siciliano) from NCI and a research Grant (A95035) from AFAR (Lei Li), as well as by grants from the Swedish Cancer Fund (2163-B97-08XAC), the Swedish Radiation Protection Institute (1027.97), and Assar Gabrielsson's Fund to Per Sunnerhagen. We thank Steve J Elledge for helpful discussions and critical review of the manuscript. The automated DNA sequencing and FACs analysis were performed in the MDACC core facilities supported by grant CA16672.
PY - 1999/3/4
Y1 - 1999/3/4
N2 - The RAD17 gene product of S. Pombe is an essential component of the checkpoint control pathway which responds to both DNA damage and disruption of replication. We have identified a human cDNA that encodes a polypeptide which is structurally conserved with the S. Pombe Rad17 protein. The human gene, designated hRAD17, predicts an encoded protein of 590 amino acids and a molecular weight of 69 kD. Amino acid sequence alignment revealed that hRad17 has 28.3% and 52.5% similarity with the S. Pombe Rad17 protein, and 21.8% identity and 45.8% similarity to the budding yeast cell cycle checkpoint protein, Rad 24. When introduced into the S. Pombe rad17 mutant, hRAD17 was able to partially revert its hydroxyurea and ionizing radiation hypersensitivity, but not its UV hypersensitivity. Permanent overexpression of the hRAD17 gene in human fibrosarcoma cells resulted in p53 activation and a significant reduction of S- and G2/M-phase cells accompanied by an accumulation of the G1-phase population, suggesting that hRAD17 may have a role in cell cycle checkpoint control. Immunostaining of HT-1080 cells transiently transfected with a hRAD17 construct confirmed the nuclear accumulation of p53, which mimics the induction caused by DNA damage. Using FISH analysis, we have mapped the hRAD17 locus to human chromosome 5q11.2.
AB - The RAD17 gene product of S. Pombe is an essential component of the checkpoint control pathway which responds to both DNA damage and disruption of replication. We have identified a human cDNA that encodes a polypeptide which is structurally conserved with the S. Pombe Rad17 protein. The human gene, designated hRAD17, predicts an encoded protein of 590 amino acids and a molecular weight of 69 kD. Amino acid sequence alignment revealed that hRad17 has 28.3% and 52.5% similarity with the S. Pombe Rad17 protein, and 21.8% identity and 45.8% similarity to the budding yeast cell cycle checkpoint protein, Rad 24. When introduced into the S. Pombe rad17 mutant, hRAD17 was able to partially revert its hydroxyurea and ionizing radiation hypersensitivity, but not its UV hypersensitivity. Permanent overexpression of the hRAD17 gene in human fibrosarcoma cells resulted in p53 activation and a significant reduction of S- and G2/M-phase cells accompanied by an accumulation of the G1-phase population, suggesting that hRAD17 may have a role in cell cycle checkpoint control. Immunostaining of HT-1080 cells transiently transfected with a hRAD17 construct confirmed the nuclear accumulation of p53, which mimics the induction caused by DNA damage. Using FISH analysis, we have mapped the hRAD17 locus to human chromosome 5q11.2.
KW - Cell cycle checkpoint
KW - DNA damage
KW - Fission yeast S. Pombe
KW - Human homolog
KW - Rad17
KW - p53
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U2 - 10.1038/sj.onc.1202469
DO - 10.1038/sj.onc.1202469
M3 - Article
C2 - 10208430
AN - SCOPUS:0033522170
SN - 0950-9232
VL - 18
SP - 1689
EP - 1699
JO - Oncogene
JF - Oncogene
IS - 9
ER -