3-36993661-C-G

Variant summary

Our verdict is Pathogenic. Variant got 18 ACMG points: 18P and 0B. PM1PM2PM5PP3_StrongPP5_Very_Strong

The NM_000249.4(MLH1):​c.114C>G​(p.Asn38Lys) variant causes a missense, splice region change involving the alteration of a non-conserved nucleotide. The variant was absent in control chromosomes in GnomAD project. In-silico tool predicts a pathogenic outcome for this variant. 2/3 splice prediction tools predict no significant impact on normal splicing. Variant has been reported in ClinVar as Pathogenic (★★★). Another nucleotide change resulting in the same amino acid substitution has been previously reported as Likely pathogenic in Lovd. Another variant affecting the same amino acid position, but resulting in a different missense (i.e. N38D) has been classified as Pathogenic.

Frequency

Genomes: not found (cov: 33)

Consequence

MLH1
NM_000249.4 missense, splice_region

Scores

15
3
1
Splicing: ADA: 0.1989
2

Clinical Significance

Pathogenic reviewed by expert panel P:5

Conservation

PhyloP100: 2.48
Variant links:
Genes affected
MLH1 (HGNC:7127): (mutL homolog 1) The protein encoded by this gene can heterodimerize with mismatch repair endonuclease PMS2 to form MutL alpha, part of the DNA mismatch repair system. When MutL alpha is bound by MutS beta and some accessory proteins, the PMS2 subunit of MutL alpha introduces a single-strand break near DNA mismatches, providing an entry point for exonuclease degradation. The encoded protein is also involved in DNA damage signaling and can heterodimerize with DNA mismatch repair protein MLH3 to form MutL gamma, which is involved in meiosis. This gene was identified as a locus frequently mutated in hereditary nonpolyposis colon cancer (HNPCC). [provided by RefSeq, Aug 2017]

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ACMG classification

Classification made for transcript

Verdict is Pathogenic. Variant got 18 ACMG points.

PM1
In a binding_site (size 0) in uniprot entity MLH1_HUMAN there are 7 pathogenic changes around while only 1 benign (88%) in NM_000249.4
PM2
Very rare variant in population databases, with high coverage;
PM5
Other missense variant is known to change same aminoacid residue: Variant chr3-36993659-A-G is described in Lovd as [Pathogenic].
PP3
MetaRNN computational evidence supports a deleterious effect, 0.995
PP5
Variant 3-36993661-C-G is Pathogenic according to our data. Variant chr3-36993661-C-G is described in ClinVar as [Pathogenic]. Clinvar id is 89651.Status of the report is reviewed_by_expert_panel, 3 stars. Variant chr3-36993661-C-G is described in Lovd as [Pathogenic].

Transcripts

RefSeq

Gene Transcript HGVSc HGVSp Effect #exon/exons MANE Protein UniProt
MLH1NM_000249.4 linkc.114C>G p.Asn38Lys missense_variant, splice_region_variant 1/19 ENST00000231790.8 NP_000240.1 P40692-1

Ensembl

Gene Transcript HGVSc HGVSp Effect #exon/exons TSL MANE Protein Appris UniProt
MLH1ENST00000231790.8 linkc.114C>G p.Asn38Lys missense_variant, splice_region_variant 1/191 NM_000249.4 ENSP00000231790.3 P40692-1

Frequencies

GnomAD3 genomes
Cov.:
33
GnomAD4 exome
Cov.:
31
GnomAD4 genome
Cov.:
33

ClinVar

Significance: Pathogenic
Submissions summary: Pathogenic:5
Revision: reviewed by expert panel
LINK: link

Submissions by phenotype

Hereditary cancer-predisposing syndrome Pathogenic:2
Likely pathogenic, criteria provided, single submitterclinical testingAmbry GeneticsJun 07, 2018The p.N38K pathogenic mutation (also known as c.114C>G), located in coding exon 1 of the MLH1 gene, results from a C to G substitution at nucleotide position 114. The asparagine at codon 38 is replaced by lysine, an amino acid with similar properties. One in vitro assay indicated that when compared to wild type MLH1, this alteration lead to a significant decrease in mismatch repair efficiency (Drost M et al. Hum. Mutat. 2010 Mar;31:247-53). Another study found that this alteration localized to the nucleus and retained interaction with PMS2 through an in vitro GST pull-down assay (Andersen SD et al. Hum. Mutat. 2012 Dec;33:1647-55). This mutation was identified in an individual with colon cancer diagnosed at 36 whose tumor exhibited microsatellite instability and copy-neutral loss of heterozygosity (van Puijenbroek M et al. Fam. Cancer 2008 Apr;7:319-30) and was detected in a patient with colon and endometrial cancers diagnosed before age 50 where the tumor tested was MSI-H and demonstrated isolated loss of PMS2 by IHC analysis (Ambry internal data). This alteration has been classified as pathogenic using the following lines of evidence: in silico prediction models, segregation with disease, clinical phenotype including tumor characteristics, mutation co-occurrence, and functional studies (Thompson BA et al. Hum. Mutat. 2013 Jan;34:200-9; Thompson BA et al. Nat. Genet. 2014 Feb;46:107-15; available at [www.insight-group.org/variants/classifications/]). This variant was not reported in population-based cohorts in the Genome Aggregation Database (gnomAD). This amino acid position is highly conserved in available vertebrate species. In addition, this alteration is predicted to be deleterious by in silico analysis. In addition, this alteration is predicted to be deleterious by MAPP-MMR in silico analyses (Chao EC et al. Hum. Mutat. 2008 Jun;29:852-60). Based on the supporting evidence, this alteration is interpreted as a disease-causing mutation. -
Likely pathogenic, criteria provided, single submitterclinical testingColor Diagnostics, LLC DBA Color HealthSep 24, 2020This missense variant replaces asparagine with lysine at codon 38 of the MLH1 protein. Computational prediction suggests that this variant may have deleterious impact on protein structure and function (internally defined REVEL score threshold >= 0.7, PMID: 27666373). Different variants affecting the same position (p.Asn38His, p.Asn38Ser, p.Asn38Thr) are considered to be disease-causing (ClinVar variation ID: 89645, 89648, 619503), suggesting that asparagine at this position is important for protein structure and function. Splice site prediction tools suggest that this variant may not impact RNA splicing. Functional studies have shown that this variant is abnormal for complementation of mismatch repair activity in MLH1-deficient HCT116 nuclear extract (PMID: 20020535) and the expression of the variant in mouse cells resulted in DNA damage tolerance (PMID: 31784484). This variant has been reported in an individual and a family affected with Lynch syndrome-associated cancer who met Bethesda and Amsterdam I criteria (PMID: 20020535, 18415027, InSiGHT database). The variant segregates with disease in the carrier family with segregation likelihood ratio of 95:1 or LOD score of approximately 2 (InSiGHT database). This variant has not been identified in the general population by the Genome Aggregation Database (gnomAD). Based on the available evidence, this variant is classified as Likely Pathogenic. -
Colorectal cancer, hereditary nonpolyposis, type 2 Pathogenic:1
Likely pathogenic, criteria provided, single submitterclinical testingMyriad Genetics, Inc.Jul 10, 2023This variant is considered likely pathogenic. Functional studies indicate this variant impacts protein function [PMID: 20020535]. This variant is expected to disrupt protein structure [Myriad internal data]. -
Lynch syndrome Pathogenic:1
Pathogenic, reviewed by expert panelresearchInternational Society for Gastrointestinal Hereditary Tumours (InSiGHT)Sep 05, 2013Multifactorial likelihood analysis posterior probability >0.99 -
Hereditary nonpolyposis colorectal neoplasms Pathogenic:1
Likely pathogenic, criteria provided, single submitterclinical testingLabcorp Genetics (formerly Invitae), LabcorpAug 22, 2022This variant is not present in population databases (gnomAD no frequency). This sequence change replaces asparagine, which is neutral and polar, with lysine, which is basic and polar, at codon 38 of the MLH1 protein (p.Asn38Lys). This missense change has been observed in individual(s) with clinical features of Lynch syndrome (PMID: 18415027). In summary, the currently available evidence indicates that the variant is pathogenic, but additional data are needed to prove that conclusively. Therefore, this variant has been classified as Likely Pathogenic. This variant disrupts the p.Asn38 amino acid residue in MLH1. Other variant(s) that disrupt this residue have been determined to be pathogenic (PMID: 12373605, 15713769, 20704743, 26895986, 27435373, 28514183). This suggests that this residue is clinically significant, and that variants that disrupt this residue are likely to be disease-causing. Algorithms developed to predict the effect of sequence changes on RNA splicing suggest that this variant may disrupt the consensus splice site. Experimental studies have shown that this missense change affects MLH1 function (PMID: 20020535, 22753075). Based on a multifactorial likelihood algorithm using genetic, in silico, and/or statistical data, this variant has been determined to have a high probability of being pathogenic (PMID: 24362816). ClinVar contains an entry for this variant (Variation ID: 89651). -

Computational scores

Source: dbNSFP v4.3

Name
Calibrated prediction
Score
Prediction
AlphaMissense
Pathogenic
1.0
BayesDel_addAF
Pathogenic
0.37
D
BayesDel_noAF
Pathogenic
0.30
CADD
Pathogenic
28
DANN
Uncertain
1.0
DEOGEN2
Pathogenic
0.97
D
Eigen
Pathogenic
0.83
Eigen_PC
Pathogenic
0.67
FATHMM_MKL
Uncertain
0.94
D
LIST_S2
Pathogenic
0.98
D
M_CAP
Pathogenic
0.84
D
MetaRNN
Pathogenic
1.0
D
MetaSVM
Uncertain
0.62
D
MutationAssessor
Pathogenic
5.0
H
PrimateAI
Pathogenic
0.82
D
PROVEAN
Pathogenic
-4.8
D
REVEL
Pathogenic
0.89
Sift
Pathogenic
0.0
D
Sift4G
Pathogenic
0.0
D
Polyphen
1.0
D
Vest4
0.98
MutPred
0.97
Gain of methylation at N38 (P = 0.0088);
MVP
0.97
MPC
0.43
ClinPred
1.0
D
GERP RS
4.2
Varity_R
0.97
gMVP
0.96

Splicing

Name
Calibrated prediction
Score
Prediction
dbscSNV1_ADA
Benign
0.20
dbscSNV1_RF
Benign
0.41
SpliceAI score (max)
0.22
Details are displayed if max score is > 0.2
DS_DL_spliceai
0.22
Position offset: 2

Find out detailed SpliceAI scores and Pangolin per-transcript scores at spliceailookup.broadinstitute.org

Publications

LitVar

Below is the list of publications found by LitVar. It may be empty.

Other links and lift over

dbSNP: rs267607706; hg19: chr3-37035152; API