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J Biol Chem, Vol. 273, Issue 29, 18547-18555, July 17, 1998

Splicing of Two Internal and Four Carboxyl-terminal Alternative Exons in Nonmuscle Tropomyosin 5 Pre-mRNA Is Independently Regulated during Development

Cécile Dufour^§, Ron P. Weinberger^§¶, Galina Schevzov^§, Peter L. Jeffrey, and Peter Gunning^§

From the  Cell Biology Unit, Children's Medical Research Institute, Wentworthville, New South Wales 2145, the ^§ Oncology Research Unit, the New Children's Hospital, P. O. Box 3515, Parramatta, New South Wales 2124, the ^¶ Developmental Neurobiology Unit, Children's Medical Research Institute, Wentworthville, New South Wales 2145, and the  Department of Paediatrics and Child Health, University of Sydney, Sydney, New South Wales 2006, Australia

Four nonmuscle tropomyosin isoforms have been reported to be produced from the rat Tm5 gene by alternative splicing (Beisel, K. W., and Kennedy, J. E. (1994) Gene (Amst.) 145, 251-256). In order to detect additional isoforms that might be expressed from that gene, we used reverse transcriptase-polymerase chain reaction assays and evaluated the presence of all product combinations of two alternative internal exons (6a and 6b) and four carboxyl-terminal exons (9a, 9b, 9c, and 9d) in developing and adult rat brain. We identified five different combinations for exon 9 (9a + 9b, 9a + 9c, 9a + 9d, 9c, and 9d), and the exon combinations 9a + 9c and 9a + 9d were previously unreported. Each of these combinations existed with both exon 6a and exon 6b. Thus, the rat brain generates at least 10 different isoforms from the Tm5 gene. Northern blot hybridization with alternative exon-specific probes revealed that these isoforms were also expressed in a number of different adult rat tissues, although some exons are preferentially expressed in particular tissues. Studies of regulation of the 10 different Tm5 isoform mRNAs during rat brain development indicated that no two isoforms are coordinately accumulated. Furthermore, there is a developmental switch in the use of exon 6a to exon 6b from embryonic to adult isoforms. TM5 protein isoforms show a differential localization in the adult cerebellum.

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