Open AccessSpecificity of RNA maturation pathways: RNAs transcribed by RNA polymerase III are not substrates for splicing or polyadenylation.
Author(s) -
Sangram S. Sisodia,
Barbara SollnerWebb,
Don W. Cleveland
Publication year - 1987
Publication title -
molecular and cellular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.14
H-Index - 327
eISSN - 1067-8824
pISSN - 0270-7306
DOI - 10.1128/mcb.7.10.3602
Subject(s) - biology , rna polymerase ii , polyadenylation , rna polymerase iii , rna polymerase i , microbiology and biotechnology , polymerase , rna splicing , small nuclear rna , transcription factor ii d , rna polymerase ii holoenzyme , transcription (linguistics) , rna dependent rna polymerase , rnase h , rna , rnase p , gene , gene expression , genetics , promoter , linguistics , philosophy
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.