HRad17 colocalizes with NHP2L1 in the nucleolus and redistributes after UV irradiation.
The rad17 gene of Schizosaccharomyces pombe plays an important role as a checkpoint protein following DNA damage and during DNA replication. The human homologue of S. pombe rad17, Hrad17, was recently identified, but its function has not yet been established. Using the yeast two-hybrid system, we determined that HRad17 can interact with a nucleolar protein, NHP2L1. This interaction was also demonstrated biochemically, in human cells. Immunofluorescence studies revealed that HRad17 and NHP2L1 colocalize to the nucleolus, and immunogold labeling further resolved the location of NHP2L1 to the dense fibrillar component of the nucleolus. Interestingly, the localization of HRad17 in the nucleolus was altered in response to UV irradiation. These results provide some insight into the DNA damage and replication checkpoint mechanisms of HRad17.
Chang
M S
MS
Department of Cancer Biology, Dana-Farber Cancer Institute and Harvard Medical School, Boston, Massachusetts 02115, USA.
Sasaki
H
H
Campbell
M S
MS
Kraeft
S K
SK
Sutherland
R
R
Yang
C Y
CY
Liu
Y
Y
Auclair
D
D
Hao
L
L
Sonoda
H
H
Ferland
L H
LH
Chen
L B
LB
D000595:Amino Acid Sequence; D018797:Cell Cycle Proteins / Q000235:genetics* / Q000378:metabolism; D002466:Cell Nucleolus / Q000235:genetics* / Q000378:metabolism; D004249:DNA Damage / Q000528:radiation effects; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal / Q000528:radiation effects*; D005800:Genes, Fungal; D006801:Humans; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D017411:Ribonucleoproteins, Small Nuclear; D029701:Saccharomyces cerevisiae Proteins; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D016415:Sequence Alignment; D014466:Ultraviolet Rays
D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D009687:Nuclear Proteins; C113575:Rad17 protein, human; D017411:Ribonucleoproteins, Small Nuclear; D029701:Saccharomyces cerevisiae Proteins; C069628:NHP2 protein, S cerevisiae