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Nucleic Acids Research, 1992, Vol. 20, No. 19 5097-5103
© 1992

MOLECULAR BIOLOGY

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

Stefan Stamm1,2, Diana Casper1, Jonathan Dinsmore3, Charles A. Kaufmann4, Jürgen Brosius1 and David M. Helfman2,*

1Fishberg Research Center in Neurobiology, The Mount Sinai School of Medicine One Gustave L.Levy Place, New York, NY 10029, USA 2Cold Spring Harbor Laboratory Cold Spring Harbor, NY 11724, USA 3Department of Biology and Center for Cancer Research, Massachusetts Institute of Technology Cambridge, MA 02139, USA 4Department of Psychiatry, Columbia University, The New York State Psychiatric Institute 722 West 168th St., New York, NY 10032, USA

* To whom correspondence should be addressed

Received June 29, 1992. Revised September 2, 1992. Accepted September 2, 1992.

The clathrin light chains are components of clathrin coated vesicles, structural constituents involved in endocytosls 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.


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