The 3'-untranslated region of membrane exon 2 from the gamma 2a immunoglobulin gene contributes to efficient transcription termination.
Elements of the mouse Immunoglobulin gamma 2a gene, near the membrane-specific poly(A) addition site, were inserted into a heterologous location in either a synthetic mouse gamma 2b gene or a gpt/SV40 chimeric gene and then assayed for their ability to terminate RNA polymerase II transcription in isolated nuclei of transfected myeloma cells. The intact gamma 2a membrane-specific 3'-untranslated region, with its potential stem loop forming sequences and poly(A) site, is able to efficiently terminate transcription in the absence of the downstream region in which transcription normally terminates (term). Termination efficiency in the presence of the termination fragment decreases either when sequences specifying a potential stem/loop, upstream of the poly(A) region, are interrupted or when the stronger membrane poly(A) site is substituted with a weaker, secretory-specific poly(A) site. We therefore conclude that the gamma 2a membrane-specific untranslated region plays a major role in specifying downstream termination. We further conclude that the immunoglobulin gamma 2a, membrane-specific, 3'-untranslated region can function in the context of the gpt gene, driven by an SV40 promoter, to terminate transcription in a poly(A) site dependent fashion.
Flaspohler
J A
JA
Department of Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pennsylvania 15261-2072, USA.
Boczkowski
D
D
Hall
B L
BL
Milcarek
C
C
D000818:Animals; D005091:Exons / Q000235:genetics*; D005803:Genes, Immunoglobulin; D005813:Genes, Synthetic; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D007074:Immunoglobulin G / Q000235:genetics*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D051379:Mice; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism; D010954:Plasmacytoma / Q000473:pathology; D011061:Poly A / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013539:Simian virus 40 / Q000235:genetics; D013728:Terminator Regions, Genetic; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D011061:Poly A; D007041:Hypoxanthine Phosphoribosyltransferase; D012319:RNA Polymerase II