Clathrin light chain B: gene structure and neuron-specific splicing

Stamm, S., Casper, D., Dinsmore, J., Kaufmann, C. A., Brosius, J., Helfman, D. M. (1992) Clathrin light chain B: gene structure and neuron-specific splicing. Nucleic Acids Res, 20 (19). pp. 5097-103. ISSN 0305-1048 (Print)0305-1048 (Linking)

URL: http://www.ncbi.nlm.nih.gov/pubmed/1408826
DOI: 10.1093/nar/20.19.5097

Abstract

The clathrin light chains are components of clathrin coated vesicles, structural constituents involved in endocytosis and membrane recycling. The clathrin light chain B (LCB) gene encodes two isoforms, termed LCB2 and LCB3, via an alternative RNA splicing mechanism. We have determined the structure of the rat clathrin light chain B gene. The gene consists of six exons that extend over 11.9 kb. The first four exons and the last exon are common to the LCB2 and LCB3 isoforms. The fifth exon, termed EN, is included in the mRNA in brain, giving rise to the brain specific form LCB2 but is excluded in other tissues, generating the LCB3 isoform. Primary rat neuronal cell cultures express predominantly the brain specific LCB2 isoform, whereas primary rat cultures of glia express only the LCB3 isoform, suggesting that expression of the brain-specific LCB2 form is limited to neurons. Further evidence for neuronal localization of the LCB2 form is provided using a teratocarcinoma cell line, P19, which can be induced by retinoic acid to express a neuronal phenotype, concomitant with the induction of the LCB2 form. In order to determine the sequences involved in alternative splice site selection, we constructed a minigene containing the alternative spliced exon EN and its flanking intron and exon sequences. This minigene reflects the splicing pattern of the endogenous gene upon transfection in HeLa cell and primary neuronal cell cultures, indicating that this region of the LCB gene contains all the necessary information for neuron-specific splicing.

Item Type: Paper
Uncontrolled Keywords: *Alternative Splicing Animals Base Sequence Brain/*physiology Cells, Cultured Clathrin/*genetics Exons HeLa Cells Humans Introns Macromolecular Substances Mesencephalon/physiology Molecular Sequence Data Neurons/*physiology Oligodeoxyribonucleotides Polymerase Chain Reaction/methods Rats Teratoma Transfection Tumor Cells, Cultured
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > Alternative Splicing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene expression
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons
organs, tissues, organelles, cell types and functions > cell types and functions > cell types > neurons
CSHL Authors:
Communities: CSHL labs > Helfman lab
Depositing User: Matt Covey
Date Deposited: 23 Sep 2015 18:38
Last Modified: 23 Sep 2015 18:38
PMCID: PMC334290
Related URLs:
URI: http://repository.cshl.edu/id/eprint/31845

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