Sequence Comparison of the Whole Murine and Human CLCA Locus Reveals Conserved Synteny Between Both Species

Ritzka, Margit; Weinel, Christian; Stanke, Frauke; Tümmler, Burkhard

doi:10.1166/153730503772757882

Cited by 8 publications

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“…Identification of mCLCA5 and mCLCA6 completes the identification of mCLCAs at the mouse CLCA locus (36). The functions of this large and diverse family are not yet fully understood, although it is clear that they have roles in the production of calcium-activated chloride currents and cell adhesion.…”

Section: Discussionmentioning

confidence: 77%

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Molecular and Functional Analyses of Two New Calcium-activated Chloride Channel Family Members from Mouse Eye and Intestine

Evans

Thoreson

Beck

2004

Journal of Biological Chemistry

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Two new calcium-activated chloride channel (CLCA) family members, mCLCA5 and mCLCA6, have been cloned from mouse eye and intestine, respectively. mCLCA5 is highly homologous to hCLCA2, and mCLCA6 is highly homologous to hCLCA4. mCLCA5 is widely expressed with strong expression in eye and spleen, whereas mCLCA6 is primarily expressed in intestine and stomach. mCLCA6 is also expressed as a splice variant lacking exon 8 and part of exon 10 in intestine and stomach. Transfection of tsA201 cells with enhanced green fluorescent protein-tagged versions of the three cDNAs reveals protein products of 155 and 65 kDa for mCLCA5 and mCLCA6 and 145 and 65 kDa for the mCLCA6 splice variant. In vitro translation of mCLCA5 generates a 90-kDa protein that does not appear to be glycosylated. mCLCA6 also generates a 90-kDa protein that is glycosylated to a 110-kDa product, whereas the mCLCA6 splice variant generates an 80-kDa product that is 100 kDa after glycosylation. Treatment of enhanced green fluorescent protein-tagged mCLCA6 with PNGase F (peptide: N-glycosidase F) to remove N-linked glycosyl groups shows a reduction in size of the 65 kDa product to 60 kDa. Consistent with the hypothesis that mCLCA5, mCLCA6, and its splice variant encode calcium-activated chloride channels, in HEK293 cells expressing CLCAs ionomycin-evoked increases in intracellular calcium stimulated a current that reversed near Cl ؊ equilibrium potential, E Cl . Furthermore, these currents were inhibited by the chloride channel blocker niflumic acid. Given the prominent role of hCLCA2 in cancer cell adhesion and the unique high level of expression of hCLCA4 in brain, the identification of their murine counterparts presents the opportunity to clarify the role of CLCAs in disease and normal cell physiology.Calcium-activated chloride currents have been characterized physiologically in many cell types including epithelium (1-4), smooth muscle (5-7), skeletal muscle (8), and neurons (9, 10). However, the molecular identities of channels responsible for calcium-activated chloride currents have not been determined.Proteins of the bestrophin and CLCA 1 families have been proposed to function as calcium-activated chloride channels (11)(12)(13)(14)(15)(16)(17)(18)(19).The CLCA gene family encodes proteins with four or five putative transmembrane domains, and many family members appear to function as calcium-activated chloride channels (bCLCA1, mCLCA1, mCLCA4, hCLCA1, and hCLCA2) (15)(16)(17)(18)(19). Other studies suggest that CLCAs may not be chloride channels themselves but may instead modulate chloride channels (20,21) In addition to their potential roles as ion channels or regulators of ion channels, some CLCA family members function as adhesion molecules. For example, bCLCA2 (Lu-ECAM-1), mCLCA1, and hCLCA2 all appear to be cell-cell adhesion molecules that help mediate the colonization of lung by lung metastatic murine B16-F10 melanoma cells (22-24). ␤ 4 integrin expressed on the surface of these melanoma cells can adhere to mCLCA1 in lung endothelium. This lea...

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Section: Discussionmentioning

confidence: 77%

“…Examination of the human and mouse CLCA loci indicates that mCLCA5 and mCLCA6 are the previously unidentified murine counterparts of hCLCA2 and hCLCA4 (36). The importance of cloning mCLCA5 and mCLCA6 is, thus, apparent in terms of the interesting roles of their human homologues.…”

mentioning

confidence: 99%

Molecular and Functional Analyses of Two New Calcium-activated Chloride Channel Family Members from Mouse Eye and Intestine

Evans

Thoreson

Beck

2004

Journal of Biological Chemistry

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“…By comparing human and mouse genomic and cDNA sequences [22] 8 loci identified in the mouse contains sequences similar to the 4 human CLCA cDNAs previously identified. These 8 mouse genes are located in a cluster on chromosome 3.…”

Section: Clca Family Expression In Adult Drgmentioning

confidence: 72%

Expression of three distinct families of calcium-activated chloride channel genes in the mouse dorsal root ganglion

et al. 2007

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Objective A calcium-activated chloride current (I Cl(Ca) ) has been observed in medium-sized sensory neurons of the dorsal root ganglion (DRG). Axotomy of the sciatic nerve induces a similar current in the majority of medium and large diameter neurons. Our aim is to identify the molecule(s) underlying this current. Methods Using conventional and quantitative RT-PCR, we examined the expression in DRG of members of three families of genes, which have been shown to have I Cl(Ca) current inducing properties. Results We showed the detection of transcripts representing several members of these families, i.e. chloride channel calciumactivated (CLCA), Bestrophin and Tweety gene families in adult DRG, in the normal state and 3 d after sciatic nerve section, a model for peripheral nerve injury. Conclusion Our analysis revealed that that mBest1 and Tweety2 appear as the best candidates to play a role in the injury-induced I Cl(Ca) in DRG neurons.

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“…On the other hand, different evolutionary paths seem to exist in different mammalian species for this gene family. For example, murine mCLCA1 and 2 have been found only in the mouse without any orthologs in the human genome (Ritzka et al 2003). Also, different functional properties have been reported for direct orthologs of different species (Gruber et al 1998a;Loewen et al 2004).…”

Section: Discussionmentioning

confidence: 99%

Murine mCLCA6 Is an Integral Apical Membrane Protein of Non-goblet Cell Enterocytes and Co-localizes With the Cystic Fibrosis Transmembrane Conductance Regulator

Bothe

Braun

Mundhenk

et al. 2008

J Histochem Cytochem.

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The CLCA family of proteins consists of a growing number of structurally and functionally diverse members with distinct expression patterns in different tissues. Several CLCA homologs have been implicated in diseases with secretory dysfunctions in the respiratory and intestinal tracts. Here we present biochemical protein characterization and details on the cellular and subcellular expression pattern of the murine mCLCA6 using specific antibodies directed against the amino- and carboxy-terminal cleavage products of mCLCA6. Computational and biochemical characterizations revealed protein processing and structural elements shared with hCLCA2 including anchorage in the apical cell membrane by a transmembrane domain in the carboxy-terminal subunit. A systematic light- and electron-microscopic immunolocalization found mCLCA6 to be associated with the microvilli of non-goblet cell enterocytes in the murine small and large intestine but in no other tissues. The expression pattern was confirmed by quantitative RT-PCR following laser-capture microdissection of relevant tissues. Confocal laser scanning microscopy colocalized the mCLCA6 protein with the cystic fibrosis transmembrane conductance regulator CFTR at the apical surface of colonic crypt cells. Together with previously published functional data, the results support a direct or indirect role of mCLCA6 in transepithelial anion conductance in the mouse intestine.

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Sequence Comparison of the Whole Murine and Human CLCA Locus Reveals Conserved Synteny Between Both Species

Cited by 8 publications

References 0 publications

Molecular and Functional Analyses of Two New Calcium-activated Chloride Channel Family Members from Mouse Eye and Intestine

Molecular and Functional Analyses of Two New Calcium-activated Chloride Channel Family Members from Mouse Eye and Intestine

Expression of three distinct families of calcium-activated chloride channel genes in the mouse dorsal root ganglion

Murine mCLCA6 Is an Integral Apical Membrane Protein of Non-goblet Cell Enterocytes and Co-localizes With the Cystic Fibrosis Transmembrane Conductance Regulator

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