Abstract
Mutations in the MTR gene, which encodes methionine synthase on human chromosome 1p43, result in the methylcobalamin deficiency G (cblG) disorder, which is characterized by homocystinuria, hyperhomocysteinemia, and hypomethioninemia. To investigate the molecular basis of the disorder, we have characterized the structure of the MTR gene, thereby identifying exon-intron boundaries. This enabled amplification of each of the 33 exons of the gene, from genomic DNA from a panel of 21 patients with cblG. Thirteen novel mutations were identified. These included five deletions (c.12-13delGC, c.381delA, c.2101delT, c.2669-2670delTG, and c.2796-2800delAAGTC) and two nonsense mutations (R585X and E1204X) that would result in synthesis of truncated proteins that lack portions critical for enzyme function. One mutation was identified that resulted in conversion of A to C of the invariant A of the 3' splice site of intron 9. Five missense mutations (A410P, S437Y, S450H, H595P, and I804T) were identified. The latter mutations, as well as the splice-site mutation, were not detected in a panel of 50 anonymous DNA samples, suggesting that these sequence changes are not polymorphisms present in the general population. In addition, a previously described missense mutation, P1173L, was detected in 16 patients in an expanded panel of 24 patients with cblG. Analysis of haplotypes constructed using sequence polymorphisms identified within the MTR gene demonstrated that this mutation, a C-->T transition in a CpG island, has occurred on at least two separate genetic backgrounds.
MeSH Terms
5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase/deficiency,genetics
Base Sequence
Chromosomes, Human, Pair 1/genetics
Codon, Nonsense
DNA Mutational Analysis
Exons
Genetic Variation
Genotype
Haplotypes
Humans
Hyperhomocysteinemia/enzymology,genetics
Introns
Molecular Sequence Data
Mutation, Missense
Phenotype
Polymorphism, Genetic
Sequence Deletion
Vitamin B 12/analogs & derivatives,metabolism
Chemicals
Codon, Nonsense
mecobalamin
5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase
Vitamin B 12
Authors & Affiliations
18 authors, click to expand affiliations / ORCID
Watkins David
Division of Medical Genetics, Department of Medicine, McGill University Health Centre, Montreal, Quebec, H3A 1A1, Canada. dwatkins@generation.net
Ru Ming
Hwang Hye-Yeon
Kim Caroline D
Murray Angus
Philip Noah S
Kim William
Legakis Helen
Wai Timothy
Hilton John F
Ge Bing
Doré Carole
Hosack Angela
Wilson Aaron
Gravel Roy A
Shane Barry
Hudson Thomas J
Rosenblatt David S
References (27)
27 references, click to expand
-
Folate and cancer prevention: a new medical application of folate beyond hyperhomocysteinemia and neural tube defects.
Nutr Rev. 1999 Oct;57(10):314-21
PMID: 10575908
-
Nonsyndromic orofacial clefts: association with maternal hyperhomocysteinemia.
Teratology. 1999 Nov;60(5):253-7
PMID: 10525201
-
Defects in methylthioadenosine phosphorylase are associated with but not responsible for methionine-dependent tumor cell growth.
Cancer Res. 2000 Oct 1;60(19):5543-7
PMID: 11034100
-
Effect of heterozygosity for the methionine synthase 2756 A-->G mutation on the risk for recurrent cardiovascular events.
Am J Cardiol. 2000 Nov 15;86(10):1144-6, A9
PMID: 11074217
-
Gene-environment and gene-gene interaction in the determination of plasma homocysteine levels in healthy middle-aged men.
Thromb Haemost. 2001 Jan;85(1):67-74
PMID: 11204591
-
Influence of a methionine synthase (D919G) polymorphism on plasma homocysteine and folate levels and relation to risk of myocardial infarction.
Atherosclerosis. 2001 Feb 15;154(3):667-72
PMID: 11257268
-
Human methionine synthase reductase, a soluble P-450 reductase-like dual flavoprotein, is sufficient for NADPH-dependent methionine synthase activation.
J Biol Chem. 2001 Sep 21;276(38):35558-63
PMID: 11466310
-
The Hordaland Homocysteine Studies.
Lipids. 2001;36 Suppl:S33-9
PMID: 11837990
-
Cloning and sequence analysis of the Escherichia coli metH gene encoding cobalamin-dependent methionine synthase and isolation of a tryptic fragment containing the cobalamin-binding domain.
J Biol Chem. 1989 Aug 15;264(23):13888-95
PMID: 2668277
-
Functional methionine synthase deficiency (cblE and cblG): clinical and biochemical heterogeneity.
Am J Med Genet. 1989 Nov;34(3):427-34
PMID: 2688421
-
Assignment of enzymatic function to specific protein regions of cobalamin-dependent methionine synthase from Escherichia coli.
Biochemistry. 1993 Sep 14;32(36):9290-5
PMID: 8369297
-
How a protein binds B12: A 3.0 A X-ray structure of B12-binding domains of methionine synthase.
Science. 1994 Dec 9;266(5191):1669-74
PMID: 7992050
-
The structure of the C-terminal domain of methionine synthase: presenting S-adenosylmethionine for reductive methylation of B12.
Structure. 1996 Nov 15;4(11):1263-75
PMID: 8939751
-
Cloning, mapping and RNA analysis of the human methionine synthase gene.
Hum Mol Genet. 1996 Dec;5(12):1851-8
PMID: 8968735
-
Defects in human methionine synthase in cblG patients.
Hum Mol Genet. 1996 Dec;5(12):1859-65
PMID: 8968736
-
Human methionine synthase: cDNA cloning and identification of mutations in patients of the cblG complementation group of folate/cobalamin disorders.
Hum Mol Genet. 1996 Dec;5(12):1867-74
PMID: 8968737
-
Human methionine synthase. cDNA cloning, gene localization, and expression.
J Biol Chem. 1997 Feb 7;272(6):3628-34
PMID: 9013615
-
Cobalamin-dependent methionine synthase is a modular protein with distinct regions for binding homocysteine, methyltetrahydrofolate, cobalamin, and adenosylmethionine.
Biochemistry. 1997 Jul 1;36(26):8082-91
PMID: 9201956
-
Cloning and mapping of a cDNA for methionine synthase reductase, a flavoprotein defective in patients with homocystinuria.
Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):3059-64
PMID: 9501215
-
Functionally null mutations in patients with the cblG-variant form of methionine synthase deficiency.
Am J Hum Genet. 1998 Aug;63(2):409-14
PMID: 9683607
-
Infant methionine synthase variants and risk for spina bifida.
J Med Genet. 1999 Jan;36(1):86-7
PMID: 9950377
-
Cloning, sequencing, and heterologous expression of rat methionine synthase cDNA.
Biosci Biotechnol Biochem. 1998 Nov;62(11):2155-60
PMID: 9972236
-
Polymorphism of the methionine synthase gene : association with homocysteine metabolism and late-onset vascular diseases in the Japanese population.
Arterioscler Thromb Vasc Biol. 1999 Feb;19(2):298-302
PMID: 9974410
-
Genetic polymorphisms in methylenetetrahydrofolate reductase and methionine synthase, folate levels in red blood cells, and risk of neural tube defects.
Am J Med Genet. 1999 May 21;84(2):151-7
PMID: 10323741
-
Relationship between total plasma homocysteine, polymorphisms of homocysteine metabolism related enzymes, risk factors and coronary artery disease in the Australian hospital-based population.
Atherosclerosis. 1999 Sep;146(1):133-40
PMID: 10487496
-
Abnormal folate metabolism and mutation in the methylenetetrahydrofolate reductase gene may be maternal risk factors for Down syndrome.
Am J Clin Nutr. 1999 Oct;70(4):495-501
PMID: 10500018
-
Recombinational and mutational hotspots within the human lipoprotein lipase gene.
Am J Hum Genet. 2000 Jan;66(1):69-83
PMID: 10631137