TY - JOUR
T1 - Specificity of RNA maturation pathways
T2 - RNAs transcribed by RNA polymerase III are not substrates for splicing or polyadenylation.
AU - Sisodia, S. S.
AU - Sollner-Webb, B.
AU - Cleveland, D. W.
PY - 1987/10
Y1 - 1987/10
N2 - To analyze the specificity of RNA processing reactions, we constructed hybrid genes containing RNA polymerase III promoters fused to sequences that are normally transcribed by polymerase II and assessed their transcripts following transfection into human 293 cells. Transcripts derived from these chimeric constructs were analyzed by using a combined RNase H and S1 nuclease assay to test whether RNAs containing consensus 5' and 3' splicing signals could be efficiently spliced in intact cells, even though they were transcribed by RNA polymerase III. We found that polymerase III-derived RNAs are not substrates for splicing. Similarly, we were not able to detect poly(A)+ RNAs derived from genes that contained a polymerase III promoter linked to sequences that were necessary and sufficient to direct 3'-end cleavage and polyadenylation when transcribed by RNA polymerase II. Our findings are consistent with the view that in vivo splicing and polyadenylation pathways are obligatorily coupled to transcription by RNA polymerase II.
AB - To analyze the specificity of RNA processing reactions, we constructed hybrid genes containing RNA polymerase III promoters fused to sequences that are normally transcribed by polymerase II and assessed their transcripts following transfection into human 293 cells. Transcripts derived from these chimeric constructs were analyzed by using a combined RNase H and S1 nuclease assay to test whether RNAs containing consensus 5' and 3' splicing signals could be efficiently spliced in intact cells, even though they were transcribed by RNA polymerase III. We found that polymerase III-derived RNAs are not substrates for splicing. Similarly, we were not able to detect poly(A)+ RNAs derived from genes that contained a polymerase III promoter linked to sequences that were necessary and sufficient to direct 3'-end cleavage and polyadenylation when transcribed by RNA polymerase II. Our findings are consistent with the view that in vivo splicing and polyadenylation pathways are obligatorily coupled to transcription by RNA polymerase II.
UR - http://www.scopus.com/inward/record.url?scp=0023424069&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0023424069&partnerID=8YFLogxK
U2 - 10.1128/MCB.7.10.3602
DO - 10.1128/MCB.7.10.3602
M3 - Article
C2 - 3683396
AN - SCOPUS:0023424069
SN - 0270-7306
VL - 7
SP - 3602
EP - 3612
JO - Molecular and cellular biology
JF - Molecular and cellular biology
IS - 10
ER -