Functional analysis of hMLH1 variants and HNPCC-related mutations using a human expression system

Trojan, Joerg; Zeuzem, Stefan; Randolph, Ann; Hemmerle, Christine; Brieger, Angela; Raedle, Jochen; Plotz, Guido; Jiricny, Josef; Marra, Giancarlo

doi:10.1053/gast.2002.30296

Cited by 190 publications

(180 citation statements)

References 48 publications

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“…The hMRE11 452-634 fragment comprises the minimal hMLH1-interacting domain, while hMRE11 1-634 (i.e., hMRE11 ATLD1) lacks the last 122 amino acid residues that include a DNA-binding motif [8]. Since 293T cells lack hMLH1 expression [4], the resulting positive clones and the parental cells were stably transfected to express hMLH1. The expression of desired proteins was validated by immunoblots (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Telephone: (509) 335-7537, FAX#: (509) 335-9688, E-mail: cher@wsu.edu. 4 These authors contributed equally to this work amino acid substitutions that are expected to produce functionally hypomorphic full-length proteins. In fact, 25% of the approximately 300 presently known hMLH1 mutations are scattered missense alterations, for which the precise roles of these mutations in the pathogenesis of Lynch syndrome are obscure [2].…”

Section: Introductionmentioning

confidence: 99%

“…To directly assess the functional effects of hMLH1 missense mutations, S. cerevisiae and human systems have been previously experimented [3,4]. In human cells, hMLH1 existed as a heterodimeric complex with hPMS2 [5], and the disruption of this heterocomplex by hMLH1 Lynch syndrome mutations, including those reside outside of the PMS2-binding domain, can lead to defective MMR [6].…”

Section: Introductionmentioning

confidence: 99%

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The interplay between hMLH1 and hMRE11: Role in MMR and the effect of hMLH1 mutations

Zhao

Zhu

Yuan

et al. 2008

Biochemical and Biophysical Research Communications

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Our previous studies indicate that hMRE11 plays a role in MMR, and this function of hMRE11 is most likely mediated by the hMLH1-hMRE11 interaction. Here, we explored the functional implications of the hMLH1-hMRE11 interaction in MMR and the effects of hMLH1 mutations on their interaction. Our in vitro MMR assay demonstrated that the dominant-negative hMRE11 452-634 mutant peptide (i.e., harboring only the hMLH1-interacting domain) imparted a significant reduction in both 3' excision and 3'-directed MMR activities. Furthermore, the expression of hMRE11 , and to a lesser extent hMRE11 1-634 (ATLD1), impaired G2/M checkpoint control in response to MNU and cisplatin treatments, rendering cells resistant to killings by these two anticancer drugs. Analysis of 38 hMLH1 missense mutations showed that the majority of mutations caused significant (> 50%) reductions in their interaction with hMRE11, suggesting a potential link between aberrant protein interaction and the pathogenic effects of hMLH1 variants.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The interplay between hMLH1 and hMRE11: Role in MMR and the effect of hMLH1 mutations

Zhao

Zhu

Yuan

et al. 2008

Biochemical and Biophysical Research Communications

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“…We found six different mutations in MSH2 and MLH1 categorised as unclassified variants; their clinical significance is uncertain. However, the Thr117Met variant in MLH1 most probably has a pathogenic effect as demonstrated in a recently developed in vitro assay (Trojan et al, 2002).…”

Section: Genetics and Genomicsmentioning

confidence: 99%

“…; (c) kindred also described by Menko et al, 1994;Wijnen et al, 1999 (NA-22); (d) pathogenicity assessed by in vitro test (Trojan et al, 2002); (e) clinical picture includes Muir -Torre syndrome; UV=unclassified variant…”

mentioning

confidence: 99%

Genomic deletions of MSH2 and MLH1 in colorectal cancer families detected by a novel mutation detection approach

et al. 2002

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Hereditary non-polyposis colorectal cancer is an autosomal dominant condition due to germline mutations in DNA-mismatchrepair genes, in particular MLH1, MSH2 and MSH6. Here we describe the application of a novel technique for the detection of genomic deletions in MLH1 and MSH2. This method, called multiplex ligation-dependent probe amplification, is a quantitative multiplex PCR approach to determine the relative copy number of each MLH1 and MSH2 exon. Mutation screening of genes was performed in 126 colorectal cancer families selected on the basis of clinical criteria and in addition, for a subset of families, the presence of microsatellite instability (MSI-high) in tumours. Thirty-eight germline mutations were detected in 37 (29.4%) of these kindreds, 31 of which have a predicted pathogenic effect. Among families with MSI-high tumours 65.7% harboured germline gene defects. Genomic deletions accounted for 54.8% of the pathogenic mutations. A complete deletion of the MLH1 gene was detected in two families. The multiplex ligation-dependent probe amplification approach is a rapid method for the detection of genomic deletions in MLH1 and MSH2. In addition, it reveals alterations that might escape detection using conventional diagnostic techniques. Multiplex ligation-dependent probe amplification might be considered as an early step in the molecular diagnosis of hereditary non-polyposis colorectal cancer.

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Molecular stool screening for colorectal cancer

Mak

Lalloo

Evans

et al. 2004

British Journal of Surgery

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Functional analysis of hMLH1 variants and HNPCC-related mutations using a human expression system

Cited by 190 publications

References 48 publications

The interplay between hMLH1 and hMRE11: Role in MMR and the effect of hMLH1 mutations

The interplay between hMLH1 and hMRE11: Role in MMR and the effect of hMLH1 mutations

Genomic deletions of MSH2 and MLH1 in colorectal cancer families detected by a novel mutation detection approach

Molecular stool screening for colorectal cancer

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