Alternative titles; symbols
HGNC Approved Gene Symbol: MSH4
Cytogenetic location: 1p31.1 Genomic coordinates (GRCh38): 1:75,796,882-75,913,242 (from NCBI)
| Location | Phenotype |
Phenotype MIM number |
Inheritance |
Phenotype mapping key |
|---|---|---|---|---|
| 1p31.1 | Premature ovarian failure 20 | 619938 | Autosomal recessive | 3 |
| Spermatogenic failure 2 | 108420 | Autosomal recessive | 3 |
The Escherichia coli MutHLS system has been highly conserved throughout evolution. The eukaryotic pathway results in a specialization of MutS homologs such as MSH2 (609309) that have evolved to play crucial roles in both DNA mismatch repair and meiotic recombination. Meiosis-specific genes belonging to this family had been identified only in yeast. In Saccharomyces cerevisiae, MSH4 (MutS homolog 4) is a meiosis-specific protein that is not involved in mismatch correction. This protein is required for reciprocal recombination and proper segregation of homologous chromosomes at meiosis I. Paquis-Flucklinger et al. (1997) identified the human MSH4 homolog. The predicted 936-amino acid sequence shows 28.7% identity with the S. cerevisiae MSH4 protein. By Northern blot analysis, human MSH4 transcripts were detectable only in testis and in ovary with a lower level of expression.
By Northern blot analysis of mouse tissues using a human MSH4 probe, Kneitz et al. (2000) detected a 3.2-kb transcript only in testis. Immunofluorescent microscopy demonstrated colocalization of mouse Msh4 with the synaptonemal complex and axial element protein Cor1 (604759) on spermatocyte meiotic chromosomes during prophase I at day 17 postpartum.
By immunohistochemistry on human testicular tissue, Tang et al. (2020) observed that MSH4 protein was highly expressed in spermatocytes.
By fluorescence in situ hybridization, Paquis-Flucklinger et al. (1997) mapped the MSH4 gene to chromosome 1p31.
Using yeast 2-hybrid analysis, Winand et al. (1998) found direct interaction between MSH4 and MSH5 (603382). Neither MSH4 nor MSH5 formed homooligomers.
To investigate whether MLH3 (604395) acts during meiotic recombination, Santucci-Darmanin et al. (2002) analyzed its expression in mammalian germ cells. The MLH3 gene was expressed in mouse meiotic cells and in human testis, and (as revealed by immunoprecipitation assays) the MLH3 protein was found in mouse spermatocytes. The meiosis-specific MSH4 protein, known to participate to meiotic recombination, coimmunoprecipitated with MLH3 from mouse meiotic cell extracts. Two MLH3 protein isoforms potentially expressed in human testis (MLH3 and MLH3-delta-7) interacted in vitro with the MSH4 protein. The authors suggested that MLH3 is associated with MSH4 in mammalian meiotic cells, and MLH3 may play a role in mammalian meiotic recombination.
Snowden et al. (2004) demonstrated that purified human MSH4-MSH5 heterodimers bound uniquely to Holliday junctions. Holliday junctions stimulated ATP hydrolysis by MSH4-MSH5 by increasing the rate of ADP-ATP exchange. The MSH4-MSH5 dimer formed a hydrolysis-independent sliding clamp that dissociated from the Holliday junction crossover region, embracing 2 homologous duplex DNA arms. Snowden et al. (2004) concluded that the MSH4-MSH5 heterodimers stabilize and preserve a meiotic bimolecular double-strand break repair intermediate.
Cannavo et al. (2020) showed that human MutS-gamma, a complex of MSH4 and MSH5 that supports crossing over, bound branched recombination intermediates and associated with MutL-gamma, a complex of MLH1 (120436) and MLH3, stabilizing the ensemble at joint molecule structures and adjacent double-stranded DNA. MutS-gamma directly stimulated DNA cleavage by the MutL-gamma endonuclease. MutL-gamma activity was further stimulated by exonuclease-1 (EXO1; 606063), but only when MutS-gamma was present. Replication factor C (RFC; see 102579) and proliferating cell nuclear antigen (PCNA; 176740) were additional components of the nuclease ensemble, thereby triggering crossing over. S. cerevisiae strains in which MutL-gamma could not interact with Pcna presented defects in forming crossovers. The MutL-gamma-MutS-gamma-EXO1-RFC-PCNA nuclease ensemble preferentially cleaved DNA with Holliday junctions, but it showed no canonical resolvase activity. Instead, the data suggested that the nuclease ensemble processed meiotic recombination intermediates by nicking double-stranded DNA adjacent to the junction points. The authors proposed that, since DNA nicking by MutL-gamma depends on its cofactors, the asymmetric distribution of MutS-gamma and RFC-PCNA on meiotic recombination intermediates may drive biased DNA cleavage. They suggested that this mode of MutL-gamma nuclease activation may explain crossover-specific processing of Holliday junctions or their precursors in meiotic chromosomes.
Independently, Kulkarni et al. (2020) showed that PCNA was important for crossover-biased resolution. In vitro assays with human enzymes showed that PCNA and RFC were sufficient to activate the MutL-gamma endonuclease. MutL-gamma was further stimulated by the codependent activity of the pro-crossover factors EXO1 and MutS-gamma, the latter of which binds Holliday junctions. The authors found that MutL-gamma also bound various branched DNAs, including Holliday junctions, but it did not show canonical resolvase activity, suggesting that the endonuclease incises adjacent to junction branch points to achieve resolution. In vivo, Rfc facilitated MutL-gamma-dependent crossing over in budding yeast. Moreover, Pcna localized to prospective crossover sites along synapsed chromosomes. Kulkarni et al. (2020) concluded that their data highlight similarities between crossover resolution and the initiation steps of DNA mismatch repair and evoke a novel model for crossover-specific resolution of double Holliday junctions during meiosis.
Premature Ovarian Failure 20
By whole-exome sequencing (WES) in 2 Colombian sisters with premature ovarian failure (POF20; 619938), Carlosama et al. (2017) identified homozygosity for a splicing mutation in the MSH4 gene (602105.0001). Sanger sequencing confirmed familial segregation, and the variant was not found in 135 ethnically and geographically matched female controls, or in the ExAC database. Exon-trapping/minigene assays demonstrated that the mutation causes skipping of exon 17, which was confirmed by Sanger sequencing.
By WES and Sanger sequencing in a consanguineous Iranian family with both male and female infertility, Akbari et al. (2021) identified homozygosity for a missense mutation in the MSH4 gene (S754L; 602105.0003) in 5 infertile sibs, including 2 sisters with POF. The first-cousin parents and 3 fertile sisters were heterozygous for the mutation, which was found at low minor allele frequency (0.0001) in the gnomAD database, but never in homozygosity.
In a Dutch woman with infertility due to premature ovarian failure and her infertile brother who was azoospermic (SPGF2; 108420), Wyrwoll et al. (2022) identified homozygosity for a nonsense mutation in the MSH4 gene (S733X; 602105.0008). Their distantly related parents were heterozygous for the variant, which was found at low minor allele frequency (0.00006087) in the gnomAD database.
Spermatogenic Failure 2
By WES in an infertile Han Chinese man with azoospermia due to meiotic arrest (SPGF2; 108420), Tang et al. (2020) identified homozygosity for a nonsense mutation in the MSH4 gene (Q518X; 602105.0002). Sanger sequencing showed that his consanguineous parents were heterozygous for the mutation, which was not found in public variant databases.
By WES and Sanger sequencing in a consanguineous Iranian family with both male and female infertility, Akbari et al. (2021) identified homozygosity for a missense mutation in the MSH4 gene (S754L; 602105.0003) in 5 infertile sibs, including 3 brothers with azoospermia. The first-cousin parents were heterozygous for the mutation, as was an infertile brother with oligozoospermia. The variant was found at low minor allele frequency (0.0001) in the gnomAD database, but never in homozygosity.
By WES and Sanger sequencing in 4 Chinese male probands with infertility due to azoospermia, Li et al. (2022) identified homozygosity or compound heterozygosity for mutations in the MSH4 gene (see, e.g., 602105.0004-602105.0007). The mutations were found in heterozygosity in the respective parents and were not found or were present at low minor allele frequency in the ExAC and gnomAD databases.
By exome sequencing in an infertile Dutch man (AMC-01) who was part of the DARWIN study and was negative for deletions in the Y-chromosome AZF region and for mutations in 133 male infertility-associated genes, Wyrwoll et al. (2022) identified homozygosity for a nonsense mutation in the MSH4 gene (S733X; 602105.0008). The proband had an infertile sister with premature ovarian failure (see POF20) who was also homozygous for the S733X mutation; their distantly related parents were heterozygous for the variant, which was found at low minor allele frequency (0.00006087) in the gnomAD database. Wyrwoll et al. (2022) also analyzed exome data from the MERGE study, consisting of 1,305 men with various infertility phenotypes, of whom 1,056 had azoospermia or cryptozoospermia, including 90 men with meiotic arrest. Chromosomal aberrations and Y-chromosome AZF microdeletions were excluded in all of the men. The authors identified a German man (M2047) who was compound heterozygous for 2 nonsense mutations (Q485X and V563X) in the MSH4 gene. The unaffected mother was heterozygous for Q485X, and a fertile brother was heterozygous for V563X; DNA was unavailable from the proband's father.
Kneitz et al. (2000) generated mice deficient in Msh4. These mice were viable and underwent normal development, apart from low testicular weight. Msh4 -/- males had normal sexual behavior and aggressiveness, but were infertile. Early after birth of Msh4 -/- females, most oocytes were lost from the ovaries, indicating a loss of oocytes before dictyate arrest. One month after birth, wildtype ovaries were oocyte-rich, whereas ovaries in Msh4 -/- females were very small and nearly devoid of oocytes. Chromosomal pairing was impaired in Msh4 -/- spermatocytes and oocytes and was associated with increased localization of Rad51 (179617) foci on meiotic chromosomes, a phenotype similar to that observed in Msh5 -/- mice. Double-mutant mice lacking both Msh4 and Msh5 were also viable but infertile. Chromosomal pairing analysis suggested that Msh5 acts in the same pathway but upstream of Msh4 due to the greater impairment in the degree of pairing.
In 2 Colombian sisters with premature ovarian failure (POF20; 619938), Carlosama et al. (2017) identified homozygosity for a donor splice site mutation in intron 17 of the MSH4 gene (c.2355+1G-A). Exon-trapping/minigene assays in HeLa cells demonstrated that the mutation causes skipping of exon 17 with deletion of 43 amino acids (Ile743_Lys785del), including the highly conserved Walker B motif of the ATP-binding domain. Sanger sequencing confirmed the mutation and its presence in heterozygosity in their unaffected parents, who came from the same isolated region of Colombia. Their fertile sister and 2 fertile brothers did not carry the variant, which was not found in the ExAC database or in 135 women from the same region of Colombia who were over 50 years of age and did not have a history of infertility.
In an infertile Han Chinese man with azoospermia due to meiotic arrest (SPGF2; 108420), Tang et al. (2020) identified homozygosity for a c.1552C-T transition in the MSH4 gene, resulting in a gln518-to-ter (Q518X) substitution. His unaffected consanguineous parents were heterozygous for the mutation, which was not found in the 1000 Genomes Project, ExAC, or gnomAD databases. Immunohistochemical analysis of a testicular biopsy showed no MSH4 expression in patient spermatocytes, and qRT-PCR showed significantly decreased levels of MSH4 mRNA in patient testicular tissue compared to control tissue.
In a consanguineous Iranian family with both male infertility due to azoospermia (SPGF2; 108420), and female infertility due to premature ovarian failure (POF20; 619938), Akbari et al. (2021) identified homozygosity for a c.2261C-T transition (c.2261C-T, NM_002440.4) in exon 17 of the MSH4 gene, resulting in a ser754-to-leu (S754L) substitution at a highly conserved residue initiating the signature motif of MutS domain V. Five infertile sibs were homozygous for the variant, including 2 sisters with POF and 3 brothers with azoospermia. Sanger sequencing confirmed the mutation and its presence in heterozygosity in the first-cousin parents and 3 fertile sisters, as well as in an infertile brother with oligozoospermia. The variant was found at low minor allele frequency (0.0001) in the gnomAD database, but never in homozygosity.
In an infertile Chinese man (family P9359) with azoospermia due to meiotic arrest at the spermatocyte stage (SPGF2; 108420), Li et al. (2022) identified homozygosity for an 8-bp deletion (c.805_812del, NM_002440.4) in the MSH4 gene, causing a frameshift predicted to result in a premature termination codon (Val269GlnfsTer15) and loss of MutS domains III to V. His parents were heterozygous for the deletion, which was not found in the ExAC or gnomAD databases.
In 2 infertile Chinese brothers (family P9517) with azoospermia due to meiotic arrest at the spermatocyte stage (SPGF2; 108420), Li et al. (2022) identified compound heterozygosity for mutations in the MSH4 gene: a c.1950G-A transition (c.1950G-A, NM_002440.4), resulting in a trp650-to-ter (W650X) substitution, and a 1-bp deletion (c.2179delG), causing a frameshift predicted to result in a premature termination codon (Asp727MetfsTer11). Their parents and a fertile sister were each heterozygous for 1 of the variants. The 1-bp deletion was not found in public variant databases, but the W650X mutation was present in the ExAC and gnomAD databases at low minor allele frequency (8.3 x 10(-6) and 1.2 x 10(-5), respectively).
For discussion of the 1-bp deletion (c.2179delG, NM_002440.4) in the MSH4 gene, causing a frameshift predicted to result in a premature termination codon (Asp727MetfsTer11), that was found in compound heterozygous state in 2 infertile Chinese brothers (family P9517) with azoospermia due to meiotic arrest at the spermatocyte stage (SPGF2; 108420) by Li et al. (2022), see 602105.0005.
In an infertile Chinese man (family P21504) with azoospermia due to meiotic arrest at the spermatocyte stage (SPGF2; 108420), Li et al. (2022) identified homozygosity for a 4-bp deletion (c.2220_2223del, NM_002440.4) in the MSH4 gene, causing a frameshift predicted to result in a premature termination codon (Lys741ArgfsTer2) and loss of MutS domain V. His parents were heterozygous for the deletion, which was not found in the ExAC or gnomAD databases.
In a Dutch woman (AMC-02) with infertility due to premature ovarian failure (POF20; 619938) and her infertile brother (AMC-01) who was azoospermic due to meiotic arrest (SPGF2; 108420), Wyrwoll et al. (2022) identified homozygosity for a c.2198C-A transversion in the MSH4 gene, resulting in a ser733-to-ter (S733X) substitution. Their distantly related parents were heterozygous for the variant, which was found at low minor allele frequency (0.00006087) in the gnomAD database.
Akbari, A., Padidar, K., Salehi, N., Mashayekhi, M., Almadani, N., Gilani, M. A. S., Bashambou, A., McElreavey, K., Totonchi, M. Rare missense variant in MSH4 associated with primary gonadal failure in both 46,XX and 46,XY individuals. Hum. Reprod. 36: 1134-1145, 2021. [PubMed: 33448284] [Full Text: https://doi.org/10.1093/humrep/deaa362]
Cannavo, E., Sanchez, A., Anand, R., Ranjha, L., Hugener, J., Adam, C., Acharya, A., Weyland, N., Aran-Guiu, X., Charbonnier, J.-B., Hoffmann, E. R., Borde, V., Matos, J., Cejka, P. Regulation of the MLH1-MLH3 endonuclease in meiosis. Nature 586: 618-622, 2020. Note: Erratum: Nature 590: E29, 2021. Electronic Article. [PubMed: 32814904] [Full Text: https://doi.org/10.1038/s41586-020-2592-2]
Carlosama, C., Elzaiat, M., Patino, L. C., Mateus, H. E., Veitia, R. A., Laissue, P. A homozygous donor splice-site mutation in the meiotic gene MSH4 causes primary ovarian insufficiency. Hum. Molec. Genet. 26: 3161-3166, 2017. [PubMed: 28541421] [Full Text: https://doi.org/10.1093/hmg/ddx199]
Kneitz, B., Cohen, P. E., Avdievich, E., Zhu, L., Kane, M. F., Hou, H., Jr., Kolodner, R. D., Kucherlapati, R., Pollard, J. W., Edelmann, W. MutS homolog 4 localization to meiotic chromosomes is required for chromosome pairing during meiosis in male and female mice. Genes Dev. 14: 1085-1097, 2000. [PubMed: 10809667]
Kulkarni, D. S., Owens, S. N., Honda, M., Ito, M., Yang, Y., Corrigan, M. W., Chen, L., Quan, A. L., Hunter, N. PCNA activates the MutL-gamma endonuclease to promote meiotic crossing over. Nature 586: 623-627, 2020. Note: Erratum: Nature 590: E30, 2021. Electronic Article. [PubMed: 32814343] [Full Text: https://doi.org/10.1038/s41586-020-2645-6]
Li, P., Ji, Z., Zhi, E., Zhang, Y., Han, S., Zhao, L., Tian, R., Chen, H., Huang, Y., Zhang, J., Chen, H., Zhao, F., Zhou, Z., Li, Z., Yao, C. Novel bi-allelic MSH4 variants causes meiotic arrest and non-obstructive azoospermia. Reprod. Biol. Endocr. 20: 21, 2022. [PubMed: 35090489] [Full Text: https://doi.org/10.1186/s12958-022-00900-x]
Paquis-Flucklinger, V., Santucci-Darmanin, S., Paul, R., Saunieres, A., Turc-Carel, C., Desnuelle, C. Cloning and expression analysis of a meiosis-specific MutS homolog: the human MSH4 gene. Genomics 44: 188-194, 1997. [PubMed: 9299235] [Full Text: https://doi.org/10.1006/geno.1997.4857]
Santucci-Darmanin, S., Neyton, S., Lespinasse, F., Saunieres, A., Gaudray, P., Paquis-Flucklinger, V. The DNA mismatch-repair MLH3 protein interacts with MSH4 in meiotic cells, supporting a role for this MutL homolog in mammalian meiotic recombination. Hum. Molec. Genet. 11: 1697-1706, 2002. [PubMed: 12095912] [Full Text: https://doi.org/10.1093/hmg/11.15.1697]
Snowden, T., Acharya, S., Butz, C., Berardini, M., Fishel, R. hMSH4-hMSH5 recognizes Holliday junctions and forms a meiosis-specific sliding clamp that embraces homologous chromosomes. Molec. Cell 15: 437-451, 2004. [PubMed: 15304223] [Full Text: https://doi.org/10.1016/j.molcel.2004.06.040]
Tang, D., Xu, C., Geng, H., Gao, Y., Cheng, H., Ni, X., He, X., Cao, Y. A novel homozygous mutation in the meiotic gene MSH4 leading to male infertility due to non-obstructive azoospermia. Am. J. Transl. Res. 12: 8185-8191, 2020. [PubMed: 33437391]
Winand, N. J., Panzer, J. A., Kolodner, R. D. Cloning and characterization of the human and Caenorhabditis elegans homologs of the Saccharomyces cerevisiae MSH5 gene. Genomics 53: 69-80, 1998. [PubMed: 9787078] [Full Text: https://doi.org/10.1006/geno.1998.5447]
Wyrwoll, M. J., van Walree, E. S., Hamer, G., Rotte, N., Motazacker, M. M., Meijers-Heijboer, H., Alders, M., Meissner, A., Kaminsky, E., Woste, M., Krallmann, C., Kliesch, S., Hunt, T. J., Clark, A. T., Silber, S., Stallmeyer, B., Friedrich, C., van Pelt, A. M. M., Mathijssen, I. B., Tuttelmann, F. Bi-allelic variants in DNA mismatch repair proteins MutS homolog MSH4 and MSH5 cause infertility in both sexes. Hum. Reprod. 37: 178-189, 2022. [PubMed: 34755185] [Full Text: https://doi.org/10.1093/humrep/deab230]