An alternative intronic promoter of the Bombyx A3 cytoplasmic actin gene exhibits a high level of transcriptional activity in mammalian cells.
Previous observations have indicated that the Bombyx mori gene for A3 cytoplasmic actin and vertebrate actin genes might make use of similar mechanisms for regulation of gene expression. To examine the suggested similarities, we have analyzed the expression of a LacZ reporter plasmid construct containing the 5' and 3' regulatory regions and the first intron of the A3 actin gene in a variety of vertebrate cell lines. We found that this silkworm expression cassette could drive expression of foreign genes in both mammalian and avian cell lines. Detailed analysis, however, indicated that neither the CArG box nor any of the promoter elements previously identified in the A3 actin gene were required for expression in mammalian cells. On the other hand, the first intron contained an efficient promoter, exhibiting in mouse cells a transcriptional activity comparable to that of the SV40 early promoter. The first intron of the A3 gene was also found to contain enhancer-like DNA elements that could stimulate the heterologous SV40 early promoter in mammalian cells. Promoter activity of the first intron of the A3 actin gene has not been observed previously. Recently however, we described a rare A3 actin mRNA isoform in B. mori cells, which initiates within the first intron. We suggest that the identified intronic promoter may be active not only in vertebrate cells but also in silkworm, and that it regulates the synthesis of the alternative A3 actin mRNA isoform. The characteristics of the 5' regulatory region of the A3 gene described here can also be exploited in the construction of bi-functional insect-mammalian expression vectors.
Fatyol
K
K
Center for Molecular Biology and Gene Therapy, School of Medicine, Loma Linda University, CA 92354, USA.
Illes
K
K
Szalay
A A
AA
D020121:5' Untranslated Regions; D000199:Actins / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000235:genetics*; D016466:CHO Cells; D019556:COS Cells; D002461:Cell Line, Transformed; D006224:Cricetinae; D018076:DNA, Complementary; D005786:Gene Expression Regulation; D017344:Genes, Insect; D006801:Humans; D007438:Introns; D008322:Mammals; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
D020121:5' Untranslated Regions; D000199:Actins; D018076:DNA, Complementary