Defective expression of the DNA mismatch repair protein, MLH1, alters G2-M cell cycle checkpoint arrest following ionizing radiation

Thomas W. Davis, Carmell Wilson Van Patten, Mark Meyers, Keith A. Kunugi, Scott Cuthill, Catherine Reznikoff, Christopher Garces, C. Richard Boland, Timothy J. Kinsella, Richard Fishel, David A. Boothman

Research output: Contribution to journalArticle

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Abstract

A role for the Mut L homologue-1 (MLH1) protein, a necessary component of DNA mismatch repair (MMR), in G2-M cell cycle checkpoint arrest after 6- thioguanine (6-TG) exposure was suggested previously. A potential role for MLH1 in G1 arrest and/or G1-S transition after damage was, however, not discounted. We report that MLH1-deficient human colon carcinoma (HCT116) cells showed decreased survival and a concomitant deficiency in G2-M cell cycle checkpoint arrest after ionizing radiation (IR) compared with genetically matched, MMR-corrected human colon carcinoma (HCT116 3-6) cells. Similar responses were noted between murine MLH1 knockout compared to wild- type primary embryonic fibroblasts. MMR-deficient HCT116 cells or embryonic fibroblasts from MLH1 knockout mice also demonstrated classic DNA damage tolerance responses after 6-TG exposure. Interestingly, an enhanced p53 protein induction response was observed in HCT116 3-6 (MLH1+) compared with HCT116 (MLH1-) cells after IR or 6-TG. Retroviral vector-mediated expression of the E6 protein did not, however, affect the enhanced G2-M cell cycle arrest observed in HCT116 3-6 compared with MLH1-deficient HCT116 cells. A role for MLH1 in G2-M cell cycle checkpoint control, without alteration in G1, after IR was also suggested by similar S-phase progression between irradiated MLH1-deficient and MLH1-proficient human or murine cells. Introduction of a nocodazole-induced G2-M block, which corrected the MLH1- mediated G2-M arrest deficiency in HCT116 cells, clearly demonstrated that HCT116 and HCT116 3-6 cells did not differ in G1 arrest or G1-S cell cycle transition after IR. Thus, our data indicate that MLH1 does not play a major role in G1 cell cycle transition or arrest. We also show that human MLH1 and MSH2 steady-state protein levels did not vary with damage or cell cycle changes caused by IR or 6-TG. MLH1-mediated G2-M cell cycle delay (caused by either MMR proofreading of DNA lesions or by a direct function of the MLH1 protein in cell cycle arrest) may be important for DNA damage detection and repair prior to chromosome segregation to eliminate carcinogenic lesions (possibly brought on by misrepair) in daughter cells.

Original languageEnglish (US)
Pages (from-to)767-778
Number of pages12
JournalCancer Research
Volume58
Issue number4
StatePublished - Feb 15 1998

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MutS DNA Mismatch-Binding Protein
G2 Phase Cell Cycle Checkpoints
HCT116 Cells
Thioguanine
Cell Cycle Checkpoints
Ionizing Radiation
DNA Mismatch Repair
Cell Cycle
Proteins
DNA Damage
Colon
Fibroblasts
Carcinoma
Nocodazole
Chromosome Segregation
S Phase
Knockout Mice
DNA Repair
Survival

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Davis, T. W., Patten, C. W. V., Meyers, M., Kunugi, K. A., Cuthill, S., Reznikoff, C., ... Boothman, D. A. (1998). Defective expression of the DNA mismatch repair protein, MLH1, alters G2-M cell cycle checkpoint arrest following ionizing radiation. Cancer Research, 58(4), 767-778.

Defective expression of the DNA mismatch repair protein, MLH1, alters G2-M cell cycle checkpoint arrest following ionizing radiation. / Davis, Thomas W.; Patten, Carmell Wilson Van; Meyers, Mark; Kunugi, Keith A.; Cuthill, Scott; Reznikoff, Catherine; Garces, Christopher; Boland, C. Richard; Kinsella, Timothy J.; Fishel, Richard; Boothman, David A.

In: Cancer Research, Vol. 58, No. 4, 15.02.1998, p. 767-778.

Research output: Contribution to journalArticle

Davis, TW, Patten, CWV, Meyers, M, Kunugi, KA, Cuthill, S, Reznikoff, C, Garces, C, Boland, CR, Kinsella, TJ, Fishel, R & Boothman, DA 1998, 'Defective expression of the DNA mismatch repair protein, MLH1, alters G2-M cell cycle checkpoint arrest following ionizing radiation', Cancer Research, vol. 58, no. 4, pp. 767-778.
Davis TW, Patten CWV, Meyers M, Kunugi KA, Cuthill S, Reznikoff C et al. Defective expression of the DNA mismatch repair protein, MLH1, alters G2-M cell cycle checkpoint arrest following ionizing radiation. Cancer Research. 1998 Feb 15;58(4):767-778.
Davis, Thomas W. ; Patten, Carmell Wilson Van ; Meyers, Mark ; Kunugi, Keith A. ; Cuthill, Scott ; Reznikoff, Catherine ; Garces, Christopher ; Boland, C. Richard ; Kinsella, Timothy J. ; Fishel, Richard ; Boothman, David A. / Defective expression of the DNA mismatch repair protein, MLH1, alters G2-M cell cycle checkpoint arrest following ionizing radiation. In: Cancer Research. 1998 ; Vol. 58, No. 4. pp. 767-778.
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abstract = "A role for the Mut L homologue-1 (MLH1) protein, a necessary component of DNA mismatch repair (MMR), in G2-M cell cycle checkpoint arrest after 6- thioguanine (6-TG) exposure was suggested previously. A potential role for MLH1 in G1 arrest and/or G1-S transition after damage was, however, not discounted. We report that MLH1-deficient human colon carcinoma (HCT116) cells showed decreased survival and a concomitant deficiency in G2-M cell cycle checkpoint arrest after ionizing radiation (IR) compared with genetically matched, MMR-corrected human colon carcinoma (HCT116 3-6) cells. Similar responses were noted between murine MLH1 knockout compared to wild- type primary embryonic fibroblasts. MMR-deficient HCT116 cells or embryonic fibroblasts from MLH1 knockout mice also demonstrated classic DNA damage tolerance responses after 6-TG exposure. Interestingly, an enhanced p53 protein induction response was observed in HCT116 3-6 (MLH1+) compared with HCT116 (MLH1-) cells after IR or 6-TG. Retroviral vector-mediated expression of the E6 protein did not, however, affect the enhanced G2-M cell cycle arrest observed in HCT116 3-6 compared with MLH1-deficient HCT116 cells. A role for MLH1 in G2-M cell cycle checkpoint control, without alteration in G1, after IR was also suggested by similar S-phase progression between irradiated MLH1-deficient and MLH1-proficient human or murine cells. Introduction of a nocodazole-induced G2-M block, which corrected the MLH1- mediated G2-M arrest deficiency in HCT116 cells, clearly demonstrated that HCT116 and HCT116 3-6 cells did not differ in G1 arrest or G1-S cell cycle transition after IR. Thus, our data indicate that MLH1 does not play a major role in G1 cell cycle transition or arrest. We also show that human MLH1 and MSH2 steady-state protein levels did not vary with damage or cell cycle changes caused by IR or 6-TG. MLH1-mediated G2-M cell cycle delay (caused by either MMR proofreading of DNA lesions or by a direct function of the MLH1 protein in cell cycle arrest) may be important for DNA damage detection and repair prior to chromosome segregation to eliminate carcinogenic lesions (possibly brought on by misrepair) in daughter cells.",
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