Phosphorylation of the cytoplasmic domain of the MUC1 mucin correlates with changes in cell-cell adhesion

Quin, Rachel J.; McGuckin, Michael A.

doi:10.1002/1097-0215(20000815)87:4<499::aid-ijc6>3.0.co;2-9

Cited by 65 publications

(54 citation statements)

References 32 publications

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“…Cell surface proteins were biotinylated by incubating cell monolayers with 0.5 mg/ml biotinamidocaproic acid 3-sulfo-N-hydroxysuccinimide (Sigma) in PBS with 0.1 mM CaCl 2 and 1 mM MgCl 2 , followed by lysis as above. MUC13 was immunoprecipitated with polyclonal antisera against peptides A (1/100 dilution), B (1/100), or C (1/400) or with antisera raised against irrelevant conjugated peptides as a control, using previously described methods (22). For removal of N-linked glycans, immunoprecipitates were denatured in 0.5% SDS, 0.1 M 2-mercaptoethanol for 5 min at 100°C, reconstituted to 0.15 M Tris, 0.16% SDS, 1.6% Nonidet P-40, pH 7.5, and 100 units/ml PNGase F (Roche) for 24 h at 32°C.…”

Section: Methodsmentioning

confidence: 99%

“…For removal of N-linked glycans, immunoprecipitates were denatured in 0.5% SDS, 0.1 M 2-mercaptoethanol for 5 min at 100°C, reconstituted to 0.15 M Tris, 0.16% SDS, 1.6% Nonidet P-40, pH 7.5, and 100 units/ml PNGase F (Roche) for 24 h at 32°C. Crude lysates, immunoprecipitated biotinylated cell surface proteins and deglycosylated immunoprecipitates were prepared in SDS-PAGE buffer containing 2% SDS, with and without 5% 2-mercaptoethanol, subjected to electrophoresis either in 4 -20% gradient or 10% resolving gels with 3% stacking gels, and electrotransferred to polyvinylidene difluoride or nitrocellulose membranes as described (22). Benchmark prestained M r markers were included on each gel (Life Technologies, Inc.).…”

Section: Methodsmentioning

confidence: 99%

“…Benchmark prestained M r markers were included on each gel (Life Technologies, Inc.). Western blotting was performed as described previously (22). Briefly, membranes were blocked with 10% skim milk powder and MUC13 was detected using polyclonal antisera against peptides A (1/400 dilution), B (1/400), or C (1/4000) followed by donkey anti-mouse horseradish peroxidase conjugate (Jackson Laboratories) and ECL detection (Amer-sham Pharmacia Biotech).…”

Section: Methodsmentioning

confidence: 99%

“…mMuc13 has previously been shown, in some IL-3 dependent cell lines, to produce two transcripts, of ϳ2.2 and 3 kb, generated by use of two distinct polyadenylation signals (22). Northern blot analyses of MUC13 in colorectal cancer cell lines and colonic tissues only revealed a single band of about 3.2 kb.…”

Section: Expression Of Muc13 In Colorectal Cancers and Colorectal Canmentioning

confidence: 96%

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MUC13, a Novel Human Cell Surface Mucin Expressed by Epithelial and Hemopoietic Cells

Williams

Wreschner

Tran

et al. 2001

Journal of Biological Chemistry

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Transmembrane mucins are glycoproteins involved in barrier function in epithelial tissues. To identify novel transmembrane mucin genes, we performed a tblastn search of the GenBank EST data bases with a serine/ threonine-rich search string, and a rodent gene expressed in bone marrow was identified. We determined the cDNA sequence of the human orthologue of this gene, MUC13, which localizes to chromosome band 3q13.3 and generates 3.2-kilobase pair transcripts encoding a 512-amino acid protein comprised of an N-terminal mucin repeat domain, three epidermal growth factor-like sequences, a SEA module, a transmembrane domain, and a cytoplasmic tail (GenBank accession no. AF286113). MUC13 mRNA is expressed most highly in the large intestine and trachea, and at moderate levels in the kidney, small intestine, appendix, and stomach. In situ hybridization in murine tissues revealed expression in intestinal epithelial and lymphoid cells. Immunohistochemistry demonstrated the human MUC13 protein on the apical membrane of both columnar and goblet cells in the gastrointestinal tract, as well as within goblet cell thecae, indicative of secretion in addition to presence on the cell surface. MUC13 is cleaved, and the ␤-subunit containing the cytoplasmic tail undergoes homodimerization. Including MUC13, there are at least five cell surface mucins expressed in the gastrointestinal tract.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Expression Of Muc13 In Colorectal Cancers and Colorectal Canmentioning

confidence: 96%

See 2 more Smart Citations

MUC13, a Novel Human Cell Surface Mucin Expressed by Epithelial and Hemopoietic Cells

Williams

Wreschner

Tran

et al. 2001

Journal of Biological Chemistry

Self Cite

280

246

View full text Add to dashboard Cite

show abstract

“…The cytoplasmic tail of MUC1 is also phosphorylated at tyrosine residue. In vitro, tyrosine phosphorylation of the MUC1 cytoplasmic domain increases recolonisation and promotes changes in cell -cell adhesion (Quin and McGuckin, 2000). Interestingly, both src and EGFR induce tyrosine phosphorylation and enhance interactions between MUC1 and b-catenin (Li et al, 2001;Schroeder et al, 2001).…”

Section: Implication Of Mucins In the Pathogenesis Of Pcmentioning

confidence: 99%

Multiple roles of mucins in pancreatic cancer, a lethal and challenging malignancy

et al. 2004

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Mucins are members of an expanding family of large multifunctional glycoproteins. Pancreatic mucins have important biological functions, including the protection, lubrication, and moisturisation of the surfaces of epithelial tissues lining ductal structures within the pancreas. Several lines of evidence support the notion that deregulated mucin production is a hallmark of inflammatory and neoplastic disorders of the pancreas. Herein, we discuss the factors that contribute to the lethality of pancreatic cancer as well as the key role played by mucins, particularly MUC1 and MUC4, in the development and progression of the disease. Aspects pertaining to the aberrant expression and glycosylation of mucins are discussed, with special emphasis on their potential impact on the design and implementation of adequate diagnostic and therapeutic strategies for combating this lethal malignancy.

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Human MUC1 mucin: a potent glandular morphogen

et al. 2001

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Human MUC1 mucin is a high-molecular-weight transmembrane glycoprotein, which is apically expressed in the majority of glandular epithelia. During embryonic development, changes in the pattern of MUC1 mucin expression coincide with the onset of glandular differentiation. This mucin is also frequently overexpressed and aberrantly glycosylated in carcinomas. To investigate the potential role of MUC1 mucin in morphogenesis, a full length MUC1 cDNA was transfected into murine mammary adenocarcinoma (410.4) and Madin-Darby canine kidney (MDCK) cells. This generated four clonal cell lines. Western blotting, FACS analysis, and immunohistochemistry confirmed expression of MUC1. All four MUC1-expressing clones demonstrated altered morphogenesis when cultured in three-dimensional type I collagen gels. While parental and vector control 410.4 cells formed compact spherical structures, the MUC1-expressing clones formed complex branching structures. Similarly, while parental and vector control MDCK cells formed small circumscribed colonies with a central lumen, the MUC1-expressing clones formed elongated tubules. MUC1 expression was also associated with reduced cellular cohesion and enhanced migration on type I collagen-coated surfaces for all except one of the clones, which expressed only low levels of MUC1 on the cell surface. These results show that MUC1 expression stimulates morphogenetic changes in two distinct epithelial cell lines. Taken together with previous observations on MUC1 expression in embryonic development and carcinomas, this finding suggests that MUC1 may induce changes in tissue architecture in both normal development and cancer.

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Phosphorylation of the cytoplasmic domain of the MUC1 mucin correlates with changes in cell-cell adhesion

Cited by 65 publications

References 32 publications

MUC13, a Novel Human Cell Surface Mucin Expressed by Epithelial and Hemopoietic Cells

MUC13, a Novel Human Cell Surface Mucin Expressed by Epithelial and Hemopoietic Cells

Multiple roles of mucins in pancreatic cancer, a lethal and challenging malignancy

Human MUC1 mucin: a potent glandular morphogen

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