TY - JOUR
T1 - Complementation group assignments of moderately UV-sensitive CHO mutants isolated by large-scale screening (FAECB)
AU - Busch, David
AU - Greiner, Carol
AU - Rosenfeld, Kathy Lewis
AU - Ford, Ruth
AU - De Wit, Jan
AU - Hoeijmakers, Jan H.J.
AU - Thompson, Larry H.
N1 - Funding Information:
For technical assistance, we thank Jerry Adams, Roberta Bliss Albeit, John Couch, Lloyd Davis, Lucille Lee, Alex Para, Donna Phillips, Lisa Wills Plater, Helen Stauffer and Sheri Stewart. This work was supported by The Armed Forces Institute of Pathology (APC=UBDH), the American Registry of Pathology and the National Institutes of Health (grant nos GM22021, CA04484 and RR00961) and performed under the auspices of the US Department of Energy by Lawrence Berkely Laboratory grant no. 7134700 and by the Lawrence Uvermore National Laboratory under contract W-74O5-ENG-48. The opinions or assertions are the private views of the authors and are not to be construed as official or as representing the views of the Department of the Army or Department of Defense. The comments on large-scale mutant isolation represent the views of D.Busch.
PY - 1994/7
Y1 - 1994/7
N2 - The complementation group (CG) assignment is presented for 74 moderately sensitive (∼2-4× sensitivity of parental line based on ratio of D10S) UV-sensitive mutants of Chinese hamster ovary (CHO) cells from the Facility for Automated Experiments in Cell Biology (FAECB) collection. The distribution of mutants within the first five rodent UV CGs was similar to that of previously reported highly sensitive (>4× wild-type UV sensitivity) mutants from this collection. This analysis nearly completes the identification of this large collection of over 200 mutant lines isolated after screening an estimated 3 million total colonies of mutagenized CHO cells from ∼20 mutant hunts with up to about 400 000 colonies screened. Only eight lines with less than about 2× parental line UV sensitivity remain unassigned. One CG of UV mutants (CG6), which now has five identified representatives in the collection, has only been found among moderately UV-sensitive CHO cells. Mutant UV40, a mitomycin C (MMC)- and X-ray-sensitive line with moderate UV cross-sensitivity, is not in CGs 1-6 and apparently is not a nucleotide excision repair mutant. Also identified were new alleles of CG1 and CG4 mutants with profoundly deficient unscheduled DNA synthesis and moderate UV sensitivity but low sensitivity to MMC. The first CG5 mutant derived from MMC-sensitive MC5 cells has been identified as the second CG5 mutant in the collection. No representatives of rodent CGs 7-11 were found, suggesting that AA8 cells have a chromosomal makeup that precludes easy isolation of mutants in these CGs. The only mutagenesis treatment generating mutants in all of the CGs 1-6 as well as the MMC mutant UV40 was with the frameshift mutagen ICR170. New CGs were not found using mutagen-sensitive derivatives of AA8 (EM9, MC5) in place of wild-type cells, although these parental cell lines did allow the isolation of new double mutant genotypes.
AB - The complementation group (CG) assignment is presented for 74 moderately sensitive (∼2-4× sensitivity of parental line based on ratio of D10S) UV-sensitive mutants of Chinese hamster ovary (CHO) cells from the Facility for Automated Experiments in Cell Biology (FAECB) collection. The distribution of mutants within the first five rodent UV CGs was similar to that of previously reported highly sensitive (>4× wild-type UV sensitivity) mutants from this collection. This analysis nearly completes the identification of this large collection of over 200 mutant lines isolated after screening an estimated 3 million total colonies of mutagenized CHO cells from ∼20 mutant hunts with up to about 400 000 colonies screened. Only eight lines with less than about 2× parental line UV sensitivity remain unassigned. One CG of UV mutants (CG6), which now has five identified representatives in the collection, has only been found among moderately UV-sensitive CHO cells. Mutant UV40, a mitomycin C (MMC)- and X-ray-sensitive line with moderate UV cross-sensitivity, is not in CGs 1-6 and apparently is not a nucleotide excision repair mutant. Also identified were new alleles of CG1 and CG4 mutants with profoundly deficient unscheduled DNA synthesis and moderate UV sensitivity but low sensitivity to MMC. The first CG5 mutant derived from MMC-sensitive MC5 cells has been identified as the second CG5 mutant in the collection. No representatives of rodent CGs 7-11 were found, suggesting that AA8 cells have a chromosomal makeup that precludes easy isolation of mutants in these CGs. The only mutagenesis treatment generating mutants in all of the CGs 1-6 as well as the MMC mutant UV40 was with the frameshift mutagen ICR170. New CGs were not found using mutagen-sensitive derivatives of AA8 (EM9, MC5) in place of wild-type cells, although these parental cell lines did allow the isolation of new double mutant genotypes.
UR - http://www.scopus.com/inward/record.url?scp=0028358137&partnerID=8YFLogxK
U2 - 10.1093/mutage/9.4.301
DO - 10.1093/mutage/9.4.301
M3 - Article
C2 - 7968570
AN - SCOPUS:0028358137
SN - 0267-8357
VL - 9
SP - 301
EP - 306
JO - Mutagenesis
JF - Mutagenesis
IS - 4
ER -