Abstract
Mucin genes encode the polypeptide backbone of the mucin glycoproteins which are expressed on all epithelial surfaces and are major constituents of the mucus layer. Mucins are, thus, expressed at the interface between the external and the internal environment of the organism, and represent the first line of defence of our body. These genes often have an extensive region of repetitive exonic sequence which codes for the heavily glycosylated domain, whose roles include bacterial interactions and gel hydration. This region shows, in several of the genes, considerable inter-individual variation in repeat number and sequence. Because of their site of expression and their high variability in this important domain, mucin genes are good candidates for conferring differences in genetic susceptibility to multifactorial epithelial and inflammatory disease. However, progress in characterizing the genes has been considerably slower than the rest of the genome because of their size and the GC-rich content of the large, repetitive variable region. Some of the issues relating to the study of these genes are discussed in this chapter. In addition, methods and approaches that have been used successfully are described.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Pigny, P., Guyonnet-Duperat, V., Hill, A. S., Pratt, W. S., Galiegue-Zouitina, S., d’Hooge, M. C., Laine, A., Van-Seuningen, I., Degand, P., Gum, J. R., Kim, Y. S., Swallow, D. M., Aubert, J. P., and Porchet, N. (1996) Human mucin genes assigned to 11p15.5: identification and organization of a cluster of genes. Genomics 38, 340–352.
Chen, Y., Zhao, Y. H., Kalaslavadi, T. B., Hamati, E., Nehrke, K., Le, A. D., Ann, D. K., and Wu, R. (2004) Genome-wide search and identification of a novel gel-forming mucin MUC19/Muc19 in glandular tissues. Am J Respir Cell Mol Biol 30, 155–165.
Thornton, D. J., Rousseau, K., and McGuckin, M. A. (2008) Structure and function of the polymeric mucins in airways mucus. Annu Rev Physiol 70, 459–486.
Swallow, D. M., Gendler, S., Griffiths, B., Corney, G., Taylor-Papadimitriou, J., and Bramwell, M. E. (1987) The human tumour-associated epithelial mucins are coded by an expressed hypervariable gene locus PUM. Nature 328, 82–84.
Vinall, L. E., King, M., Novelli, M., Green, C. A., Daniels, G., Hilkens, J., Sarner, M., and Swallow, D. M. (2002) Altered expression and allelic association of the hypervariable membrane mucin MUC1 in Helicobacter pylori gastritis. Gastroenterology 123, 41–49.
Silva, F., Carvalho, F., Peixoto, A., Seixas, M., Almeida, R., Carneiro, F., Mesquita, P., Figueiredo, C., Nogueira, C., Swallow, D. M., Amorim, A., and David, L. (2001) MUC1 gene polymorphism in the gastric carcinogenesis pathway. Eur J Hum Genet 9, 548–552.
Silva, F., Carvalho, F., Peixoto, A., Teixeira, A., Almeida, R., Reis, C., Bravo, L. E., Realpe, L., Correa, P., and David, L. (2003) MUC1 polymorphism confers increased risk for intestinal metaplasia in a Colombian population with chronic gastritis. Eur J Hum Genet 11, 380–384.
Toribara, N. W., Gum, J. R., Jr., Culhane, P. J., Lagace, R. E., Hicks, J. W., Petersen, G. M., and Kim, Y. S. (1991) MUC-2 human small intestinal mucin gene structure. Repeated arrays and polymorphism. J Clin Invest 88, 1005–1013.
van de Bovenkamp, J. H., Hau, C. M., Strous, G. J., Buller, H. A., Dekker, J., and Einerhand, A. W. (1998) Molecular cloning of human gastric mucin MUC5AC reveals conserved cysteine-rich D-domains and a putative leucine zipper motif. Biochem Biophys Res Commun 245, 853–859.
Escande, F., Aubert, J. P., Porchet, N., and Buisine, M. P. (2001) Human mucin gene MUC5AC: organization of its 5’-region and central repetitive region. Biochem J 358, 763–772.
Engelmann, K., Baldus, S. E., and Hanisch, F. G. (2001) Identification and topology of variant sequences within individual repeat domains of the human epithelial tumor mucin MUC1. J Biol Chem 276, 27764–27769.
Fowler, J. C., Teixeira, A. S., Vinall, L. E., and Swallow, D. M. (2003) Hypervariability of the membrane-associated mucin and cancer marker MUC1. Hum Genet 113, 473–479.
von Mensdorff-Pouilly, S., Kinarsky, L., Engelmann, K., Baldus, S. E., Verheijen, R. H., Hollingsworth, M. A., Pisarev, V., Sherman, S., and Hanisch, F. G. (2005) Sequence-variant repeats of MUC1 show higher conformational flexibility, are less densely O-glycosylated and induce differential B lymphocyte responses. Glycobiology 15, 735–746.
Ng, W., Loh, A. X., Teixeira, A. S., Pereira, S. P., and Swallow, D. M. (2008) Genetic regulation of MUC1 alternative splicing in human tissues. Br J Cancer.
Kamio, K., Matsushita, I., Hijikata, M., Kobashi, Y., Tanaka, G., Nakata, K., Ishida, T., Tokunaga, K., Taguchi, Y., Homma, S., Nakata, K., Azuma, A., Kudoh, S., and Keicho, N. (2005) Promoter analysis and aberrant expression of the MUC5B gene in diffuse panbronchiolitis. Am J Respir Crit Care Med 171, 949–957.
Loh, A. X., Johnson, L., Ng, W., and Swallow, D. M. Cis-acting allelic variation in MUC5B mRNA expression is associated with different promoter haplotypes. Ann Hum Genet 74, 498–505.
Rousseau, K., Vinall, L. E., Butterworth, S. L., Hardy, R. J., Holloway, J., Wadsworth, M. E., and Swallow, D. M. (2006) MUC7 haplotype analysis: results from a longitudinal birth cohort support protective effect of the MUC7*5 allele on respiratory function. Ann Hum Genet 70, 417–427.
Rousseau, K., Byrne, C., Griesinger, G., Leung, A., Chung, A., Hill, A. S., and Swallow, D. M. (2007) Allelic association and recombination hotspots in the mucin gene (MUC) complex on chromosome 11p15.5. Ann Hum Genet 71, 561–569.
Pigman, W., Moschera, J., Weiss, M., and Tettamanti, G. (1973) The occurrence of repetitive glycopeptide sequences in bovine submaxillary glycoprotein. Eur J Biochem 32, 148–154.
Tettamanti, G., and Pigman, W. (1968) Purification and characterization of bovine and ovine submaxillary mucins. Arch Biochem Biophys 124, 41–50.
Blair, G. W., Folley, S. J., Malpress, F. H., and Coppen, F. M. (1941) Variations in certain properties of bovine cervical mucus during the oestrous cycle. Biochem J 35, 1039–1049.
Escande, F., Porchet, N., Aubert, J. P., and Buisine, M. P. (2002) The mouse Muc5b mucin gene: cDNA and genomic structures, chromosomal localization and expression. Biochem J 363, 589–598.
Escande, F., Porchet, N., Bernigaud, A., Petitprez, D., Aubert, J. P., and Buisine, M. P. (2004) The mouse secreted gel-forming mucin gene cluster. Biochim Biophys Acta 1676, 240–250.
Chen, Y., Zhao, Y. H., and Wu, R. In silico cloning of mouse Muc5b gene and upregulation of its expression in mouse asthma model. Am J Respir Crit Care Med 164, 1059–1066.
Bhargava, A. K., Woitach, J. T., Davidson, E. A., and Bhavanandan, V. P. (1990) Cloning and cDNA sequence of a bovine submaxillary gland mucin-like protein containing two distinct domains. Proc Natl Acad Sci USA 87, 6798–6802.
Jiang, W., Gupta, D., Gallagher, D., Davis, S., and Bhavanandan, V. P. (2000) The central domain of bovine submaxillary mucin consists of over 50 tandem repeats of 329 amino acids. Chromosomal localization of the BSM1 gene and relations to ovine and porcine counterparts. Eur J Biochem 267, 2208–2217.
Jiang, W., Woitach, J. T., Gupta, D., and Bhavanandan, V. P. (1998) Sequence of a second gene encoding bovine submaxillary mucin: implication for mucin heterogeneity and cloning. Biochem Biophys Res Commun 251, 550–556.
Jiang, W., Woitach, J. T., Keil, R. L., and Bhavanandan, V. P. (1998) Bovine submaxillary mucin contains multiple domains and tandemly repeated non-identical sequences. Biochem J 331 (Pt 1), 193–199.
Lang, T., Hansson, G. C., and Samuelsson, T. (2007) Gel-forming mucins appeared early in metazoan evolution. Proc Natl Acad Sci USA 104, 16209–16214.
Lang, T., Hansson, G. C., and Samuelsson, T. (2006) An inventory of mucin genes in the chicken genome shows that the mucin domain of Muc13 is encoded by multiple exons and that ovomucin is part of a locus of related gel-forming mucins. BMC Genomics 7, 197.
Gum, J. R., Byrd, J. C., Hicks, J. W., Toribara, N. W., Lamport, D. T., and Kim, Y. S. (1989) Molecular cloning of human intestinal mucin cDNAs. Sequence analysis and evidence for genetic polymorphism. J Biol Chem 264, 6480–6487.
Guyonnet Duperat, V., Audie, J. P., Debailleul, V., Laine, A., Buisine, M. P., Galiegue-Zouitina, S., Pigny, P., Degand, P., Aubert, J. P., and Porchet, N. (1995) Characterization of the human mucin gene MUC5AC: a consensus cysteine-rich domain for 11p15 mucin genes? Biochem J 305 (Pt 1), 211–219.
Toribara, N. W., Roberton, A. M., Ho, S. B., Kuo, W. L., Gum, E., Hicks, J. W., Gum, J. R., Jr., Byrd, J. C., Siddiki, B., and Kim, Y. S. (1993) Human gastric mucin. Identification of a unique species by expression cloning. J Biol Chem 268, 5879–5885.
Dufosse, J., Porchet, N., Audie, J. P., Guyonnet Duperat, V., Laine, A., Van-Seuningen, I., Marrakchi, S., Degand, P., and Aubert, J. P. (1993) Degenerate 87-base-pair tandem repeats create hydrophilic/hydrophobic alternating domains in human mucin peptides mapped to 11p15. Biochem J 293 (Pt 2), 329–337.
Newton, C. R., Graham, A., Heptinstall, L. E., Powell, S. J., Summers, C., Kalsheker, N., Smith, J. C., and Markham, A. F. (1989) Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS). Nucleic Acids Res 17, 2503–2516.
Ye, S., Humphries, S., and Green, F. (1992) Allele specific amplification by tetra-primer PCR. Nucleic Acids Res 20, 1152.
Strachan, T., Read, A. (2010) Human Molecular Genetics, 4th ed., Garland Science.
Spielman, R. S., McGinnis, R. E., and Ewens, W. J. (1993) Transmission test for linkage disequilibrium: the insulin gene region and insulin-dependent diabetes mellitus (IDDM). Am J Hum Genet 52, 506–516.
Offner, G. D., Nunes, D. P., Keates, A. C., Afdhal, N. H., and Troxler, R. F. (1998) The amino-terminal sequence of MUC5B contains conserved multifunctional D domains: implications for tissue-specific mucin functions. Biochem Biophys Res Commun 251, 350–355.
Heazlewood, C. K., Cook, M. C., Eri, R., Price, G. R., Tauro, S. B., Taupin, D., Thornton, D. J., Png, C. W., Crockford, T. L., Cornall, R. J., Adams, R., Kato, M., Nelms, K. A., Hong, N. A., Florin, T. H., Goodnow, C. C., and McGuckin, M. A. (2008) Aberrant mucin assembly in mice causes endoplasmic reticulum stress and spontaneous inflammation resembling ulcerative colitis. PLoS Med 5, e54.
Griffiths, B., Matthews, D. J., West, L., Attwood, J., Povey, S., Swallow, D. M., Gum, J. R., and Kim, Y. S. (1990) Assignment of the polymorphic intestinal mucin gene (MUC2) to chromosome 11p15. Ann Hum Genet 54, 277–285.
Pigny, P., Pratt, W. S., Laine, A., Leclercq, A., Swallow, D. M., Nguyen, V. C., Aubert, J. P., and Porchet, N. (1995) The MUC5AC gene: RFLP analysis with the Jer58 probe. Hum Genet 96, 367–368.
Vinall, L. E., Hill, A. S., Pigny, P., Pratt, W. S., Toribara, N., Gum, J. R., Kim, Y. S., Porchet, N., Aubert, J. P., and Swallow, D. M. (1998) Variable number tandem repeat polymorphism of the mucin genes located in the complex on 11p15.5. Hum Genet 102, 357–366.
Rousseau, K., Byrne, C., Kim, Y. S., Gum, J. R., Swallow, D. M., and Toribara, N. W. (2004) The complete genomic organization of the human MUC6 and MUC2 mucin genes. Genomics 83, 936–939.
Gum, J. R., Jr., Hicks, J. W., Toribara, N. W., Rothe, E. M., Lagace, R. E., and Kim, Y. S. The human MUC2 intestinal mucin has cysteine-rich subdomains located both upstream and downstream of its central repetitive region. J Biol Chem 267, 21375–21383.
Zhu, L., Lee, P., Yu, D., Tao, S., and Chen, Y. (2010) Cloning and Characterization of Human MUC19 Gene. Am J Respir Cell Mol Biol In Press.
Jia, Y., Persson, C., Hou, L., Zheng, Z., Yeager, M., Lissowska, J., Chanock, S. J., Chow, W. H., and Ye, W. (2010) A comprehensive analysis of common genetic variation in MUC1, MUC5AC, MUC6 genes and risk of stomach cancer. Cancer Causes Control 21, 313–321.
Ubell, M. L., Khampang, P., and Kerschner, J. E. (2009) Mucin gene polymorphisms in otitis media patients. Laryngoscope 120, 132–138.
Kwon, J. A., Lee, S. Y., Ahn, E. K., Seol, S. Y., Kim, M. C., Kim, S. J., Kim, S. I., Chu, I. S., and Leem, S. H. (2010) Short rare MUC6 minisatellites-5 alleles influence susceptibility to gastric carcinoma by regulating gene. Hum Mutat 31, 942–949.
Garcia, E., Carvalho, F., Amorim, A., and David, L. (1997) MUC6 gene polymorphism in healthy individuals and in gastric cancer patients from northern Portugal. Cancer Epidemiol Biomarkers Prev 6, 1071–1074.
Nguyen, T. V., Janssen, M., Jr., Gritters, P., te Morsche, R. H., Drenth, J. P., van Asten, H., Laheij, R. J., and Jansen, J. B. (2006) Short mucin 6 alleles are associated with H pylori infection. World J Gastroenterol 12, 6021–6025.
Ahn, E. K., Kim, W. J., Kwon, J. A., Choi, P. J., Kim, W. J., Sunwoo, Y., Heo, J., and Leem, S. H. (2009) Variants of MUC5B minisatellites and the susceptibility of bladder cancer. DNA Cell Biol 28, 169–176.
Vinall, L. E., Fowler, J. C., Jones, A. L., Kirkbride, H. J., de Bolos, C., Laine, A., Porchet, N., Gum, J. R., Kim, Y. S., Moss, F. M., Mitchell, D. M., and Swallow, D. M. (2000) Polymorphism of human mucin genes in chest disease: possible significance of MUC2. Am J Respir Cell Mol Biol 23, 678–686.
Jeong, Y. H., Kim, M. C., Ahn, E. K., Seol, S. Y., Do, E. J., Choi, H. J., Chu, I. S., Kim, W. J., Kim, W. J., Sunwoo, Y., and Leem, S. H. (2007) Rare exonic minisatellite alleles in MUC2 influence susceptibility to gastric carcinoma. PLoS ONE 2, e1163.
Moehle, C., Ackermann, N., Langmann, T., Aslanidis, C., Kel, A., Kel-Margoulis, O., Schmitz-Madry, A., Zahn, A., Stremmel, W., and Schmitz, G. (2006) Aberrant intestinal expression and allelic variants of mucin genes associated with inflammatory bowel disease. J Mol Med 84, 1055–1066.
Swallow, D. M., Vinall, L. E., Gum, J. R., Kim, Y. S., Yang, H., Rotter, J. I., Mirza, M., Lee, J. C., and Lennard-Jones, J. E. (1999) Ulcerative colitis is not associated with differences in MUC2 mucin allele length. J Med Genet 36, 859–860.
Chuang, S. C., Juo, S. H., Hsi, E., Wang, S. N., Tsai, P. C., Yu, M. L., and Lee, K. T. (2011) Multiple mucin genes polymorphisms are associated with gallstone disease in Chinese men. Clin Chim Acta 412, 599–603.
Barrett, J. C., Hansoul, S., Nicolae, D. L., Cho, J. H., Duerr, R. H., Rioux, J. D., Brant, S. R., Silverberg, M. S., Taylor, K. D., Barmada, M. M., Bitton, A., Dassopoulos, T., Datta, L. W., Green, T., Griffiths, A. M., Kistner, E. O., Murtha, M. T., Regueiro, M. D., Rotter, J. I., Schumm, L. P., Steinhart, A. H., Targan, S. R., Xavier, R. J., Libioulle, C., Sandor, C., Lathrop, M., Belaiche, J., Dewit, O., Gut, I., Heath, S., Laukens, D., Mni, M., Rutgeerts, P., Van Gossum, A., Zelenika, D., Franchimont, D., Hugot, J. P., de Vos, M., Vermeire, S., Louis, E., Cardon, L. R., Anderson, C. A., Drummond, H., Nimmo, E., Ahmad, T., Prescott, N. J., Onnie, C. M., Fisher, S. A., Marchini, J., Ghori, J., Bumpstead, S., Gwilliam, R., Tremelling, M., Deloukas, P., Mansfield, J., Jewell, D., Satsangi, J., Mathew, C. G., Parkes, M., Georges, M., and Daly, M. J. (2008) Genome-wide association defines more than 30 distinct susceptibility loci for Crohn’s disease. Nat Genet 40, 955–962.
Guo, X., Pace, R. G., Stonebraker, J. R., Commander, C. W., Dang, A. T., Drumm, M. L., Harris, A., Zou, F., Swallow, D. M., Wright, F. A., O’Neal, W. K., and Knowles, M. R. (2011) Mucin variable number tandem repeat polymorphisms and severity of cystic fibrosis lung disease: significant association with MUC5AC. PLoS One 6(10), e25452.
Seibold, M. A., Wise, A. L., Speer, M. C., Steele, M. P., Brown, K. K., Loyd, J. E., Fingerlin, T. E., Zhang, W., Gudmundsson, G., Groshong, S. D., Evans, C. M., Garantziotis, S., Adler, K. B., Dickey, B. F., du Bois, R. M., Yang, I. V., Herron, A., Kervitsky, D., Talbert, J. L., Markin, C., Park. J., Crews, A. L., Slifer, S. H., Auerbach, S., Roy, M. G., Lin, J., Hennessy, C. E., Schwarz, M. I., and Schwartz, D. A. (2011) A common MUC5B promoter polymorphism and pulmonary fibrosis. Engl J Med 364(16), 1503–1512.
Vinall LE, Pratt WS, Swallow DM. Detection of mucin gene polymorphism. Methods Mol Biol. 2000;125:337–50(16), 1503–1512.
Guo X, Pace RG, Stonebraker JR, Commander CW, Dang AT, Drumm ML, Harris A, Zou F, Swallow DM, Wright FA, O’Neal WK, Knowles MR. Mucin variable number tandem repeat polymorphisms and severity of cystic fibrosis lung disease: significant association with MUC5AC. PLoS One. 2011;6(10):e25452. Epub 2011 Oct 6.
Seibold MA, Wise AL, Speer MC, Steele MP, Brown KK, Loyd JE, Fingerlin TE, Zhang W, Gudmundsson G, Groshong SD, Evans CM, Garantziotis S, Adler KB, Dickey BF, du Bois RM, Yang IV, Herron A, Kervitsky D, Talbert JL, Markin C, Park J, Crews AL, Slifer SH, Auerbach S, Roy MG, Lin J, Hennessy CE, Schwarz MI, Schwartz DA.A common MUC5B promoter polymorphism and pulmonary fibrosis. N Engl J Med, 2011 Apr 21;364(16):1503–12.
Acknowledgements
The authors would like to thank Lynne Vinall (61), Lauren Johnson and Ralph Burgess whose work (Johnson PhD thesis UCL 2010; Burgess summer project, 2006) helped in the assembly of the information described in this chapter. KR was funded by the Horserace Betting Levy Board and the Medical Research Council.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Rousseau, K., Swallow, D.M. (2012). Mucin Methods: Genes Encoding Mucins and Their Genetic Variation with a Focus on Gel-Forming Mucins. In: McGuckin, M., Thornton, D. (eds) Mucins. Methods in Molecular Biology, vol 842. Humana Press. https://doi.org/10.1007/978-1-61779-513-8_1
Download citation
DOI: https://doi.org/10.1007/978-1-61779-513-8_1
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-512-1
Online ISBN: 978-1-61779-513-8
eBook Packages: Springer Protocols