title: MUC1 splice variants in human ocular surface tissues: possible differences between dry eye patients and normal controls.
abstract: Mucins are highly glycosylated proteins that are vital to the maintenance of healthy epithelial surfaces including the ocular surface. Mucins act as lubricants, protectants, and mediators of signal transduction. The majority of the O-glycosylation sites on the transmembrane mucin MUC1 are found in a highly polymorphic core region containing a variable number of tandem repeats (VNTR). MUC1 alleles can be divided into size classes that contain small (30-45) or large (60-90) numbers of repeats. Although at least 12 splice variants of MUC1 have been found in other tissues, no splice variants have been reported in human ocular surface tissues. We have used RT-PCR to identify MUC1 splice variants that were then confirmed by sequencing. We here report the presence in some samples of human cornea, conjunctiva, and lacrimal gland of MUC1/B which features canonical splicing between exons 1 and 2 and MUC1/A, a transcript that retains 27bp from the 3' end of intron 1 and is predicted to add 9 amino acids to the MUC1 sequence upstream of the tandem repeat region. Cornea and conjunctiva both contain the MUC1/SEC splice variant that lacks the transmembrane domain and, therefore, results in a soluble, secreted form of MUC1. Cornea and conjunctiva also contain MUC1/Y and MUC1/Z(X) variants that lack the tandem repeat region. Cornea, conjunctiva, and lacrimal gland also contain a previously undescribed MUC1 variant transcript, termed MUC1/YI, that retains 99bp from the 5' end and 27bp from the 3' end of the first intron, resulting in a frame shift and premature stop codon. This transcript is predicted to produce a novel 27 amino acid peptide after signal peptidase cleavage. Analysis of brush cytology samples revealed that the percentage of dry eye patients expressing the MUC1/A variant in the conjunctival epithelium is lower than in normal control donors. Western blotting confirmed that MUC1/A is associated with alleles containing the large size class of tandem repeats. Therefore, we propose that one factor in susceptibility to dry eye disease may be the lengths of the MUC1 VNTR in conjunctival epithelium based on the rationale that longer VNTR provide better lubrication and greater protection of the ocular surface against inflammation.
authors.affiliation: Department of Molecular, Cellular & Craniofacial Biology, School of Dentistry, University of Louisville, 501 S. Preston St., Louisville, KY 40292, USA.
authors.forename: Yoannis
authors.initials: Y
authors.lastname: Imbert
authors.forename: Douglas S
authors.initials: DS
authors.lastname: Darling
authors.forename: Marcia M
authors.initials: MM
authors.lastname: Jumblatt
authors.forename: Gary N
authors.initials: GN
authors.lastname: Foulks
authors.forename: Erica G
authors.initials: EG
authors.lastname: Couzin
authors.forename: Pamela S
authors.initials: PS
authors.lastname: Steele
authors.forename: William W
authors.initials: WW
authors.lastname: Young
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D016022:Case-Control Studies; D003228:Conjunctiva / Q000378:metabolism*; D015352:Dry Eye Syndromes / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D018396:Mucin-1 / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020080:Tandem Repeat Sequences; D013666:Tears / Q000737:chemistry
chemical_list: D018396:Mucin-1