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Resources » Paper

Mathews E et al. (1995) International C. elegans Meeting "MOLECULAR CLONING AND GENOMIC MAPPING OF A CALCIUM CHANNEL ALPHA 1 SUBUNIT."

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  • Comments on Mathews E et al. (1995) International C. elegans Meeting "MOLECULAR CLONING AND GENOMIC MAPPING OF A CALCIUM CHANNEL ALPHA 1 SUBUNIT." (0)

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    Status:
    Publication type:
    Meeting_abstract
    WormBase ID:
    WBPaper00021934

    Mathews E, & Snutch TP (1995). MOLECULAR CLONING AND GENOMIC MAPPING OF A CALCIUM CHANNEL ALPHA 1 SUBUNIT presented in International C. elegans Meeting. Unpublished information; cite only with author permission.

    We have previously identified and cloned a family of five distinct types of voltage-gated calcium channels expressed in the mammalian central nervous system (Snutch and Reiner, Current Opinion Neurobiol. 2:247-253; Soong et al, Science 260:1133-1136). We used degenerate primers homologous to cloned neuronal alpha1 subunits and the polymerase chain reaction to amplify calcium channel sequences from C. elegans total RNA. The resulting 327 bp fragment hybridized to three overlapping YACs (Y76F7, Y23A3 and Y5E3). Subsequently, eight cosmids corresponding to these YACs were screened and the PCR fragment (pCe2) localized to a single cosmid (T02C5). These experiments localized a calcium channel alpha1 subunit gene to the left arm of the X chromosome near unc-2. Approximately 6 kb of sequence has been determined from TO2C5 identifying an ancestral alpha1 subunit that shares considerable amino acid identity with rat brain neuronal alpha1 subunits (41 to 65%). Northern blot analysis using staged N2 RNA reveals a single ~ 7.5 kb transcript expressed throughout development. High stringency genomic DNA and YAC blot analyses suggest that pCe2 represents a single copy element in the worm genome and does not cross-hybridize with loci previously identified as calcium channel alpha1 subunit genes (egl-19 on LG IV, Lobel et al, WBG 13(2):71 and YAC Y24H1 on LG II, Lobel et al, WBG 13(1):46). These results suggest that C. elegans possesses a diverse family of voltage-gated calcium channels. unc-2 has recently been identified as a calcium channel alpha1 subunit involved in adaptation to dopamine (W. Schafer and C. Kenyon WBG 13(3):79). Using a precomplementation screen, we have isolated eleven new alleles of unc-2 and are currently determining the sequence alterations of these mutants.


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