Restricted accessResearch articleFirst published online 2010-08
The Ethnospecific Distribution of the HFE Haplotypes for IVS2(+4)t/c,IVS4(−44)t/c,and IVS5(−47)g/a in Populations of Russia and Possible Effects of These Single-Nucleotide Polymorphisms in Splicing
Aim: The aim of this work was a haplotype analysis of the major mutations (C282Y, H63D, S65C) and IVS2(+4)t/c, IVS4(−44)t/c, and IVS5(−47)a/g polymorphisms of the hemochromatosis HFE gene in populations inhabiting the territories of Russia (Russians, Finno-Ugrians, Central Asians, and Arctic Mongoloids). Method: The hemochromatosis gene (HFE) alleles were detected using the polymerase chain reaction/restriction fragment length polymorphism method. Results: Of the eight possible intronic haplotype variants, the TTG, TTA, CTA, and CCA were identified. The HFE alleles with the different haplotype variants were distributed in an ethnospecific manner among the populations. Our finding was that every one of the C282Y, H63D, and S65C mutations was in linkage disequilibrium only with one of the intronic haplotype variants: TTG, CTA, and CCA, respectively. The data from context analysis of DNA regions where the examined single-nucleotide polymorphisms are located suggested their involvement in splicing. Conclusions: Different genotypes of the HFE gene occur at different frequencies among populations of Russia. Carriers of the specific genotype variants may potentially express distinct sets of alternative HFE mRNAs.
Get full access to this article
View all access options for this article.
References
1.
AhmadKA, AhmannJR, MigasMCet al.2002. Decreased liver hepcidin expression in the Hfe knockout mouse. Blood Cells Mol Dis, 29:361-366.
2.
AjiokaRS, KushnerJP. 2003. Clinical consequences of iron overload in hemochromatosis homozygotes. Blood, 101:3351-3354.
3.
BastinJ, DrakesmithH, ReesMet al.2006. Localisation of proteins of iron metabolism in the human placenta and liver. Br J Haematol, 134:532-543.
4.
BeutlerE. 1997. The significance of the 187G (H63D) mutation in hemochromatosis. Am J Hum Genet, 61:762-764.
5.
BeutlerE. 2003. The HFE Cys282Tyr mutation as a necessary but not sufficient cause of clinical hereditary hemochromatosis. Blood, 101:3347-3350.
6.
BeutlerE, FelittiVJ, KoziolJAet al.2002. Penetrance of 845G->A (C282Y) HFE hereditary haemochromatosis mutation in the USA. Lancet, 359:211-218.
BeutlerE, WestC. 1997. New diallelic markers in the HLA region of chromosome 6. Blood Cells Mol Dis, 23:219-229.
9.
BridleKR, FrazerDM, WilkinsSJet al.2003. Disrupted hepcidin regulation in HFE-associated haemochromatosis and the liver as a regulator of body iron homeostasis. Lancet, 361:669-673.
10.
CarmelI, TalS, VigI, AstG. 2004. Comparative analysis detects dependencies among the 5′ splice-site positions. RNA, 10:828-840.
11.
DaviesPS, EnnsCA. 2004. Expression of the hereditary hemochromatosis protein HFE increases ferritin levels by inhibiting iron export in HT29 cells. J Biol Chem, 279:25085-25092.
12.
De LucasAP, FulgencioMG, RoblesJMet al.2005. Is the IVS2 + 4 T/C variant of the HFE gene a splicing mutation or a polymorphism? A study in the Spanish population. Genet Med, 7:212-213.
13.
De VilliersJNP, HillermannR, LoubserL, KotzeMJ. 1999. Spectrum of mutations in the HFE gene implicated in haemochromatosis and porphyria. Hum Mol Genet, 8:1517-1522.
14.
DrakesmithH, SweetlandE, SchimanskiLet al.2002. The hemochromatosis protein HFE inhibits iron export from macrophages. Proc Natl Acad Sci USA, 99:15602-15607.
15.
FederJN, GnirkeA, ThomasWet al.1996. A novel MHC class I-like gene is mutated in patients with hereditary hemochromatosis. Nat Genet, 13:399-408.
16.
FederJN, TsuchihashiZ, IrrinkiAet al.1997. The hemochromatosis founder mutation in HLA-H disrupts β2-microglobulin interaction and cell surface expression. J Biol Chem, 272:14025-14028.
FlemingRE, BrittonRS. 2006. Iron imports. VI HFE and regulation of intestinal iron absorption. Am J Physiol Gastrointest Liver Physiol, 290:G590-G594.
19.
FloreaniA, NavagliaF, BassoDet al.2005. Intron 2 [IVS2, T-C + 4] HFE gene mutation associated with S65C causes alternative RNA splicing and is responsible for iron overload. Hepatol Res, 33:57-60.
20.
FujiiT, IshitaniA, GeraghtyDE. 1994. A soluble form of the HLA-G antigen is encoded by a messenger ribonucleic acid containing intron 4. J Immunol, 153:5516-5524.
21.
GalantePAF, SakabeNJ, Kirschbaum-SlagerN, de SouzaSJ. 2004. Detection and evaluation of intron retention events in the human transcriptome. RNA, 10:757-765.
22.
GoswamiT, AndrewsNC. 2006. Hereditary hemochromatosis protein, HFE, interaction with transferrin receptor 2 suggests a molecular mechanism for mammalian iron sensing. J Biol Chem, 281:28494-28498.
23.
GriffithsWJH, CoxTM. 2003. Co-localization of the mammalian hemochromatosis gene product (HFE) and a newly identified transferrin receptor (TfR2) in intestinal tissue and cells. J Histochem Cytochem, 51:613-623.
24.
GrossCN, IrrinkiA, FederJN, EnnsCA. 1998. Co-trafficking of HFE, a nonclassical major histocompatibility complex class I protein, with the transferrin receptor implies a role in intracellular iron regulation. J Biol Chem, 273:22068-22074.
25.
GuthrieC, PattersonB. 1988. Spliceosomal snRNAs. Annu Rev Genet, 22:387-419.
26.
HansonEH, ImperatoreG, BurkeW. 2001. HFE gene and hereditary hemochromatosis: a HuGe review. Am J Epidemiol, 154:193-206.
27.
HolmströmP, MarmurJ, EggertsenGet al.2002. Mild iron overload in patients carrying the HFE S65C gene mutation: a retrospective study in patients with suspected iron overload and healthy controls. Gut, 51:723-730.
28.
IshitaniA, GeraghtyDE. 1992. Alternative splicing of HLA-G transcripts yields proteins with primary structures resembling both class I and class II antigens. Proc Natl Acad Sci USA, 89:3947-3951.
29.
JacksonHA, CarterK, DarkeCet al.2001. HFE mutations, iron deficiency and overload in 10,500 blood donors. Br J Haematol, 114:474-484.
30.
JeffreyGP, BasclainK, HajekJet al.1999. Alternate splicing produces a soluble form of the hereditary hemochromatosis protein Hfe. Blood Cells Mol Dis, 25:61-67.
31.
KirszenbaumM, MoreauP, GluckmanEet al.1994. An alternatively spliced form of HLA-G mRNA in human trophoblasts and evidence for the presence of HLA-G transcript in adult lymphocytes. Proc Natl Acad Sci USA, 91:4209-4213.
32.
LebronJA, BennettMJ, VaughnDEet al.1998. Crystal structure of the hemochromatosis protein HFE and characterization of its interaction with transferrin receptor. Cell, 93:111-123.
33.
LieuPT, HeiskalaM, PetersonPAet al.2001. The roles of iron in health and disease. Mol Asp Med, 22:1-87.
34.
LudwiczekS, TheurlI, Artner-DworzakEet al.2004. Duodenal HFE expression and hepcidin levels determine body iron homeostasis: modulation by genetic diversity and dietary iron availability. J Mol Med, 82:373-382.
35.
LundM, KjemsJ. 2002. Defining a 5′ splice site by functional selection in the presence and absence of U1 snRNA 5′ end. RNA, 8:166-179.
36.
MakuiH, SoaresRJ, JiangWet al.2005. Contribution of Hfe expression in macrophages to the regulation of hepatic hepcidin levels and iron loading. Blood, 106:2189-2195.
37.
McCulloughAJ, BergetSM. 1997. G triplets located throughout a class of small vertebrate introns enforce intron borders and regulate splice site selection. Mol Cell Biol, 17:4562-4571.
38.
McCuneCA, Al-JaderLN, HayesAMSLet al.2002. Hereditary haemochromatosis: only 1% of adult HFE C282Y homozygotes in South Wales have a clinical diagnosis of iron overload. Hum Genet, 111:538-543.
39.
Merryweather-ClarkeAT, PointonJJ, JouanolleANet al.2000. Geography of HFE C282Y and H63D mutations. Genet Test, 4:183-198.
40.
MikhailovaSV, OnopchenkoOV, SukhanovAVet al.2006. Haplotypic analysis for the HFE gene in Russian populations and in patients with common diseases. Herald Vavilov Soc Genet Breed Sci, 10:504-513(In Russian).
41.
MontosiG, PagliaP, GarutiCet al.2000. Wild-type HFE protein normalizes transferrin iron accumulation in macrophages from subjects with hereditary hemochromatosis. Blood, 96:1125-1129.
42.
MuraC, RaguenesO, FerecC. 1999. HFE mutations analysis in 711 hemochromatosis probands: evidence for S65C implication in mild form of hemochromatosis. Blood, 93:2502-2505.
43.
NagaiK, MutoY, Pomeranz KrummelDAet al.2001. Structure and assembly of the spliceosomal snRNPs. Biochem Soc Trans, 29:15-26.
44.
NicolasG, BennounM, DevauxIet al.2001. Lack of hepcidin gene expression and severe tissue iron overload in upstream stimulatory factor 2 (USF2) knockout mice. Proc Natl Acad Sci USA, 98:8780-8785.
45.
ParkkilaS, WaheedA, BrittonRSet al.1997a. Association of the transferrin receptor in human placenta with HFE, the protein defective in hereditary hemochromatosis. Proc Natl Acad Sci USA, 94:13198-13202.
46.
ParkkilaS, WaheedA, BrittonRSet al.1997b. Immunohistochemistry of HLA-H, the protein defective in patients with hereditary hemochromatosis, reveals unique pattern of expression in gastrointestinal tract. Proc Natl Acad Sci USA, 94:2534-2539.
PotekhinaES, LavrovAV, SamokhodskayaLMet al.2005. Unique genetic profile of hereditary hemochromatosis in Russians: high frequency of C282Y mutation in population, but not in patients. Blood Cells Mol Dis, 35:182-188.
49.
RhodesDA, TrowsdaleJ. 1999. Alternate splice variants of the hemochromatosis gene Hfe. Immunogenetics, 49:357-359.
50.
RochetteJ, PointonJJ, FisherCAet al.1999. Multicentric origin of hemochromatosis gene (HFE) mutations. Am J Hum Genet, 64:1056-1062.
51.
RogozinIB, MilanesiL. 1997. Analysis of donor splice sites in different eukaryotic organisms. J Mol Evol, 45:50-59.
52.
Salter-CidL, PetersonPA, YangY. 2000. The major histocompatibility complex-encoded HFE in iron homeostasis and immune function. Immunol Res, 22:43-59.
53.
SambrookJ, FritschEF, ManiatisT. 1989. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press: Cold Spring Harbor, NY.
54.
SánchezM, BrugueraM, RodésJ, OlivaR. 2001. Complete characterization of the 3′ region of the human and mouse hereditary hemochromatosis HFE gene and detection of novel splicing forms. Blood Cells Mol Dis, 27:35-43.
55.
ShawerSM, VyasJM, RodgersJR, RichRR. 1994. Antigen presentation by major histocompatibility complex class 1-β molecules. Annu Rev Immunol, 12:839-880.
56.
SorekR, AstG. 2003. Intronic sequences flanking alternatively spliced exons are conserved between human and mouse. Genome Res, 13:1631-1637.
57.
SteinerM, OcranK, GenschelJet al.2002. A homozygous HFE gene splice site mutation (IVS5 + 1 G/A) in a hereditary hemochromatosis patient of Vietnamese origin. Gastroenterology, 122:789-795.
ToomajianC, KreitmanM. 2002. Sequence variation and haplotype structure at the human HFE locus. Genetics, 161:1609-1623.
60.
VogtTM, BlackwellAD, GiannettiAMet al.2003. Heterotypic interactions between transferrin receptor and transferrin receptor 2. Blood, 10:2008-2014.
61.
WaheedA, GrubbJH, ZhouXYet al.2002. Regulation of transferrin-mediated iron uptake by HFE, the protein defective in hereditary hemochromatosis. Proc Natl Acad Sci USA, 99:3117-3122.
62.
WaheedA, ParkkilaS, ZhouXYet al.1997. Hereditary hemochromatosis: effects of C282Y and H63D mutations on association with β2-microglobulin, intracellular processing, and cell surface expression of the HFE protein in COS-7 cells. Proc Natl Acad Sci USA, 94:12384-12389.
63.
YeoGW, Van NostrandEL, LiangTY. 2007. Discovery and analysis of evolutionarily conserved intronic splicing regulatory elements. PLoS Genet, 3:0814-0829.
64.
ZhangA-S, XiongS, TsukamotoH, EnnsCA. 2004. Localization of iron metabolism- related mRNAs in rat liver indicate that HFE is expressed predominantly in hepatocytes. Blood, 103:1509-1514.
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
