L. Mertens et al., SEQUENCE AND SPATIAL REQUIREMENTS FOR REGULATED MUSCLE-SPECIFIC PROCESSING OF THE SARCOPLASMIC ENDOPLASMIC RETICULUM CA2-ATPASE 2 GENE TRANSCRIPT(), The Journal of biological chemistry, 270(18), 1995, pp. 11004-11011
Expression of the muscle-specific 2a isoform of the sarco/endoplasmic
reticulum Ca2+ ATPase (SERCA2) requires activation of an otherwise ine
fficient splicing process at the 3'-end of the primary gene transcript
, The sequence and topology requirements for this regulated splicing e
vent were studied in the BC(3)H1 myogenic cell line using a minigene c
ontaining the 3'-end of the SERCA2 gene, In undifferentiated BC(3)H1 c
ells, the splice process is made inefficient by the presence of a weak
muscle-type 5'-donor site (5'D1) and a long terminal intron, Both opt
imizing the 5'D1 and decreasing the length of the muscle-specific intr
on, induced muscle type splicing in undifferentiated myogenic cells, M
oreover, the induction of muscle-type transcripts was only observed wh
en two competing processing sites, the polyadenylation site (pA(u)) us
ed in non-muscle cells and the second neuronal 5'-donor site (5'D2), w
ere weak, Indeed, making 5'D2 consensus induced neuronal-type splicing
in undifferentiated myocytes and prevented the appearance of muscle-t
ype transcripts, Similarly, replacing the polyadenylation site (pA(u))
with a strong site almost completely inhibited muscle-type splicing a
fter myogenic differentiation. We conclude that weak processing sites
and a long terminal intron are required for tissue-dependent mRNA proc
essing of the SERCA2 transcript.