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A number of human homologues of yeast cell cycle checkpoint control genes have been identified recently. In this study, the sequence alterations in six of such novel human genes (hRAD1, hRAD9, hRAD17, hHUS1, CHK1 and CHES1) were analyzed by PCR-single-strand conformational polymorphism (PCR-SSCP) method on a panel of 25 human tumor cell lines in an attempt to search for possible in vivo cases where any of the checkpoint-related genes are altered in human systems. For hRAD9, hHUS1 or CHK1, no SSCP variant was detected in any of the cell lines tested, indicating a high stability of these genes in human cancer. Most of the SSCP variants found in the other three genes were due to single nucleotide base substitutions. Two cell lines were found to be homozygous for missense-type base substitutions, i.e., Saos-2 was homoallelic for 1637T → G in hRAD17; and COLO320DM for 1189G → A in CHES1, indicating a possible use of these cell lines for further study. The former nucleotide change in hRAD17, which causes a change of amino acid from arginine to lysine at codon 546, was supposed to be polymorphic. Considering that lysine, but not arginine, is the amino acid that is well conserved among fission yeast, mouse and monkey at the corresponding position, coexistence of both alleles in human may have a functional or selectional implication.

A human-mouse hybrid containing a human 11q22-23 fragment including the ATM locus was used to examine its capability to correct the cellular defect of ataxia-telangiectasia (A-T). Examination of 21 A-T-derived hybrids indicated that the acquired radioresistance was observed in the clones where the 11q22-23 fragment was transferred intact, but not in those where donor-derived 11q segment was lost. In one exceptional clone, the ATM locus was deleted from the transferred fragment, while it was still partially radioresistant. This partially radioresistant clone was found to include the mouse-derived fragment containing the Atm gene, the mouse homologue of human ATM gene. Similar association of partial radioresistance with the presence of mouse Atm gene was observed in three additional hybrids. The results indicate that the cellular A-T defect can be corrected by the mouse subchromosomal fragment containing the Atm gene as well as by the human 11q22-23 fragment containing the ATM gene, but apparently to a lesser extent in the former.

A human-mouse monochromosomal hybrid that contains a human t(X;11) translocated chromosome carrying pter--q23 segment of chromosome 11 was used to construct a panel of radiation-reduced hybrids. The hypoxhanthine phosphoribosyltransferase (HPRT) gene located close to the translocation breakpoint was used as a marker to select for the hybrids that preferentially retain the 11q22-23 region. Twenty-three HAT-resistant hybrids were isolated and screened by polymerase chain reaction (PCR) for the retention of 31 loci on 11q22-23 region. Among the 14 hybrids that had breakpoints within the 11q22-23 region, 6 hybrids contained fragments that extend either from centromere or telomere to the 5-Mb region spanned by GRIA4 and FDX, carrying various breakpoints within the region. This subpanel could be a potential resource to analyze the ataxia-telangiectasia disease locus and its neighboring region.