Ovarian- and Somatic-specific Transcripts of the Mosquito Clathrin Heavy Chain Gene Generated by Alternative 5′-Exon Splicing and Polyadenylation

Insect oocytes are extraordinarily specialized for receptor-mediated endocytosis of yolk protein precursors. The clathrin heavy chain (CHC) is the major structural protein of coated vesicles, the principal organelles of receptor-mediated endocytosis. To understand the role of clathrin in the development of the oocyte's powerful endocytotic machinery we determined the structure of the mosquito chc gene. The gene spans approximately 45 kilobases and its coding region is divided into seven exons, five of which encode the protein. Three distinct mature transcripts of this gene were identified in mosquito tissues. Two of them code isoforms of the CHC polypeptide differing in their NH2-terminal sequences, and are specifically expressed in female germ-line cells. The third transcript has a 3'-untranslated region about 1 kilobase longer than the other variants, and is found only in the somatic cells. Tissue-specific 5'-exon splicing and alternative polyadenylation of the primary transcript combine to give rise to these mRNAs. We identified two alternative promoters, distal and proximal, separated by approximately 10 kilobases involved in tissue-specific regulation of mosquito chc gene expression. Our data provide the first molecular evidence for complex structure and regulation of a chc gene, in this case occurring at both the transcriptional and post-transcriptional levels.