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J. Biol. Chem., Vol. 266, Issue 13, 8408-8415, May, 1991
LO Goodwin, JP Lees-Miller, MA Leonard, SB Cheley and DM Helfman
cDNA clones encoding four rat tropomyosin isoforms, termed TM-2, TM-3,
TM-5a, and TM-5b, were isolated and characterized. All are derived from the
alpha-tropomyosin gene via alternative RNA processing and the use of two
alternate promoters. The cDNA sequences predict that TM-2 and TM- 3 both
contain 284 amino acids and differ from each other only at an internal
region of the protein from amino acids 189 through 213, due to alternative
splicing of exons 6a and 6b. TM-5a and TM-5b both contain 248 amino acids
and differ from each other only at an internal exon encoding amino acids
153 through 177, also due to alternative splicing of exons 6a and 6b. The
differences in the amino acid sequence encoded by these alternate exons
affects the theoretical actin-binding pattern of the tropomyosins, such
that TM-5b is expected to bind actin with greater affinity than TM-5a. TM-2
and TM-3 are transcribed from the upstream promoter, and TM-5a and TM-5b
are transcribed from an internal promoter. In addition, all four isoforms
contain the identical COOH- terminal coding region. RNA protection analyses
revealed that the mRNA for each isoform is expressed in a number of
different tissues and cell types, although the expression of some isoforms
is restricted to particular cell types. Furthermore, the expression of mRNA
encoding these isoforms was found to be altered in a number of different
virally transformed cell lines. The changes in the expression of
tropomyosin mRNAs in transformed cells reflect changes in the relative use
of the two promoters, as well as the relative use of alternatively spliced
exons 6a and 6b.
Four fibroblast tropomyosin isoforms are expressed from the rat alpha- tropomyosin gene via alternative RNA splicing and the use of two promoters
Cold Spring Harbor Laboratory, New York 11724.
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