NM_000249.4:c.588+5G>A

Variant summary

Our verdict is Pathogenic. Variant got 10 ACMG points: 10P and 0B. PM2PP5_Very_Strong

The NM_000249.4(MLH1):​c.588+5G>A variant causes a splice region, intron change involving the alteration of a non-conserved nucleotide. The variant was absent in control chromosomes in GnomAD project. In-silico tool predicts a benign outcome for this variant. 3/3 splice prediction tools predicting alterations to normal splicing. Variant has been reported in ClinVar as Pathogenic (★★★).

Frequency

Genomes: not found (cov: 32)

Consequence

MLH1
NM_000249.4 splice_region, intron

Scores

2
Splicing: ADA: 0.9994
2

Clinical Significance

Pathogenic reviewed by expert panel P:10

Conservation

PhyloP100: 3.14
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 10 ACMG points.

PM2
Very rare variant in population databases, with high coverage;
PP5
Variant 3-37011867-G-A is Pathogenic according to our data. Variant chr3-37011867-G-A is described in ClinVar as [Pathogenic]. Clinvar id is 90285.Status of the report is reviewed_by_expert_panel, 3 stars. Variant chr3-37011867-G-A is described in Lovd as [Likely_pathogenic]. Variant chr3-37011867-G-A is described in Lovd as [Pathogenic].

Transcripts

RefSeq

Gene Transcript HGVSc HGVSp Effect Exon rank MANE Protein UniProt
MLH1NM_000249.4 linkc.588+5G>A splice_region_variant, intron_variant Intron 7 of 18 ENST00000231790.8 NP_000240.1 P40692-1

Ensembl

Gene Transcript HGVSc HGVSp Effect Exon rank TSL MANE Protein Appris UniProt
MLH1ENST00000231790.8 linkc.588+5G>A splice_region_variant, intron_variant Intron 7 of 18 1 NM_000249.4 ENSP00000231790.3 P40692-1

Frequencies

GnomAD3 genomes
Cov.:
32
GnomAD4 exome
Cov.:
30
GnomAD4 genome
Cov.:
32

ClinVar

Significance: Pathogenic
Submissions summary: Pathogenic:10
Revision: reviewed by expert panel
LINK: link

Submissions by phenotype

Lynch syndrome Pathogenic:2
Sep 05, 2013
International Society for Gastrointestinal Hereditary Tumours (InSiGHT)
Significance: Pathogenic
Review Status: reviewed by expert panel
Collection Method: research

Variant causes splicing aberrations: full inactivation of variant allele -

Mar 18, 2014
Department of Pathology and Laboratory Medicine, Sinai Health System
Significance: Pathogenic
Review Status: criteria provided, single submitter
Collection Method: clinical testing

- -

not provided Pathogenic:2
Jul 01, 2023
GeneDx
Significance: Pathogenic
Review Status: criteria provided, single submitter
Collection Method: clinical testing

Intronic +5 splice variant in a gene for which loss-of-function is a known mechanism of disease, and skipping of exon 7 has been observed (Pagenstecher et al., 2006; Peterson et al., 2013; Pinero et al., 2020); Observed in multiple individuals with suspected Lynch syndrome, including those with Lynch related tumors found to be MSI-H and/or show loss of MLH1 via immunohistochemistry (Casey et al., 2005; Wolf et al., 2005; Pagenstecher et al., 2006; Batte et al., 2014; Sunga et al., 2017); Not observed at significant frequency in large population cohorts (gnomAD); This variant is associated with the following publications: (PMID: 25525159, 16341550, 15713769, 15926618, 24933100, 28449805, 18561205, 24090359, 29887214, 30787465, 32363481) -

Oct 20, 2021
Quest Diagnostics Nichols Institute San Juan Capistrano
Significance: Pathogenic
Review Status: criteria provided, single submitter
Collection Method: clinical testing

This variant has not been reported in large, multi-ethnic general populations (http://gnomad.broadinstitute.org). In the published literature, the variant has been reported in HNPCC families (PMIDs: 18389388 (2008), 16206289 (2006), 15926618 (2005)). The variant has demonstrated to affect normal MLH1 mRNA splicing, causing a skip of exon 7 (PMIDs: 24090359 (2013), 18561205 (2008), 16341550 (2006)). Based on the available information, this variant is classified as pathogenic. -

Hereditary cancer-predisposing syndrome Pathogenic:2
May 04, 2019
Color Diagnostics, LLC DBA Color Health
Significance: Pathogenic
Review Status: criteria provided, single submitter
Collection Method: clinical testing

- -

May 06, 2022
Ambry Genetics
Significance: Pathogenic
Review Status: criteria provided, single submitter
Collection Method: clinical testing

The c.588+5G>A intronic pathogenic mutation results from a G to A substitution 5 nucleotides after coding exon 7 in the MLH1 gene. This intronic mutation has been detected in multiple families meeting Amsterdam and/or Bethesda criteria for HNPCC/Lynch syndrome, with tumor results revealing absence of MLH1 protein expression on immunohistochemistry (Ambry internal data, Casey et al. JAMA. 2005. 293(7): 799–809; Wolf et al. Int J Cancer. 2006. 118(6):1465-70; Pagenstecher et al. Hum Genet. 2006. 119: 9–22; Sunga AY et al. Cancer Genet 2017 04;212-213:1-7). Functional studies have demonstrated aberrant splicing, leading to skipping of exon 7 and a truncated protein product (Pagenstecher et al. Hum Genet. 2006. 119: 9–22; Petersen SM et al. BMC Medical Genetics 2013,14:103; Ambry internal data). This variant is considered to be rare based on population cohorts in the Genome Aggregation Database (gnomAD). This nucleotide position is highly conserved in available vertebrate species. In silico splice site analysis predicts that this alteration will weaken the native splice donor site. In addition to the clinical data presented in the literature, alterations that disrupt the canonical splice site are expected to cause aberrant splicing, resulting in an abnormal protein or a transcript that is subject to nonsense-mediated mRNA decay. As such, this alteration is classified as a disease-causing mutation. -

Colorectal cancer, hereditary nonpolyposis, type 2 Pathogenic:1
Jul 17, 2023
Myriad Genetics, Inc.
Significance: Pathogenic
Review Status: criteria provided, single submitter
Collection Method: clinical testing

This variant is considered pathogenic. mRNA analysis has demonstrated abnormal mRNA splicing occurs [PMID: 18561205]. -

Hereditary nonpolyposis colon cancer Pathogenic:1
Aug 21, 2020
Women's Health and Genetics/Laboratory Corporation of America, LabCorp
Significance: Pathogenic
Review Status: criteria provided, single submitter
Collection Method: clinical testing

Variant summary: MLH1 c.588+5G>A alters a conserved nucleotide located close to a canonical splice site and therefore could affect mRNA splicing, leading to a significantly altered protein sequence. Several computational tools predict a significant impact on normal splicing: Two predict the variant weakens a 5' splicing donor site and two predict it abolishes it. Experimental evidence from multiple studies supports these predictions demonstrating the variant affects mRNA splicing leading to exon skipping (Casey_2005, Pagenstecher_2006, Tournier_2008, Petersen_2013). The variant was absent in 251328 control chromosomes (gnomAD). c.588+5G>A has been reported in the literature in individuals affected with Hereditary Nonpolyposis Colorectal Cancer (e.g. Casey_2005, Pagenstecher_2006, Wolf_2006, Tournier_2008, Petersen_2013). These data indicate that the variant is likely to be associated with disease. Six ClinVar submitters (evaluation after 2014) cite the variant as pathogenic. Based on the evidence outlined above, the variant was classified as pathogenic. -

Muir-Torré syndrome;C1333991:Colorectal cancer, hereditary nonpolyposis, type 2;C5399763:Mismatch repair cancer syndrome 1 Pathogenic:1
Mar 13, 2024
Fulgent Genetics, Fulgent Genetics
Significance: Pathogenic
Review Status: criteria provided, single submitter
Collection Method: clinical testing

- -

Hereditary nonpolyposis colorectal neoplasms Pathogenic:1
Dec 16, 2024
Labcorp Genetics (formerly Invitae), Labcorp
Significance: Pathogenic
Review Status: criteria provided, single submitter
Collection Method: clinical testing

This sequence change falls in intron 7 of the MLH1 gene. It does not directly change the encoded amino acid sequence of the MLH1 protein. RNA analysis indicates that this variant induces altered splicing and may result in an absent or altered protein product. This variant is not present in population databases (gnomAD no frequency). This variant has been observed in individuals with Lynch syndrome (PMID: 15713769, 16341550, 18561205, 24090359, 26437257; internal data). It has also been observed to segregate with disease in related individuals. Invitae Evidence Modeling of clinical and family history, age, sex, and reported ancestry of multiple individuals with this MLH1 variant has been performed. This variant is expected to be pathogenic with a positive predictive value of at least 99%. This is a validated machine learning model that incorporates the clinical features of 1,370,736 individuals referred to our laboratory for MLH1 testing. This variant is also known as IVS07+5G>A. ClinVar contains an entry for this variant (Variation ID: 90285). Variants that disrupt the consensus splice site are a relatively common cause of aberrant splicing (PMID: 17576681, 9536098). Studies have shown that this variant results in skipping of exon 7, and produces a non-functional protein and/or introduces a premature termination codon (PMID: 15713769, 16341550, 18561205; internal data). For these reasons, this variant has been classified as Pathogenic. -

Computational scores

Source: dbNSFP v4.3

Name
Calibrated prediction
Score
Prediction
BayesDel_noAF
Benign
-0.18
CADD
Uncertain
25
DANN
Benign
0.94
RBP_binding_hub_radar
0.0
RBP_regulation_power_radar
1.1

Splicing

Name
Calibrated prediction
Score
Prediction
dbscSNV1_ADA
Pathogenic
1.0
dbscSNV1_RF
Pathogenic
0.98
SpliceAI score (max)
0.73
Details are displayed if max score is > 0.2
DS_DL_spliceai
0.73
Position offset: -5

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: rs267607768; hg19: chr3-37053358; COSMIC: COSV51622932; COSMIC: COSV51622932; API