Sequences upstream of AAUAAA influence poly(A) site selection in a complex transcription unit

The adenovirus major late transcription unit (MLTU) encodes five colinear mRNA families, L1 through L5, each distinguished by a unique poly(A) site. Site selection is regulated during the course of infection, predominating early at the L1 site and late at the L2 through L5 sites. Two general mechani...

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Vydané v:Molecular and cellular biology Ročník 9; číslo 11; s. 4951
Hlavní autori: DeZazzo, J D, Imperiale, M J
Médium: Journal Article
Jazyk:English
Vydavateľské údaje: United States 01.11.1989
Predmet:
Adenoviridae - genetics
Base Sequence
Biological Transport
Cell Line
DNA Mutational Analysis
Genetic Linkage
Humans
Plasmids
Poly A - metabolism
Promoter Regions, Genetic
Restriction Mapping
RNA, Messenger
Transcription, Genetic
ISSN:0270-7306
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Abstract The adenovirus major late transcription unit (MLTU) encodes five colinear mRNA families, L1 through L5, each distinguished by a unique poly(A) site. Site selection is regulated during the course of infection, predominating early at the L1 site and late at the L2 through L5 sites. Two general mechanisms can be invoked to explain predominant usage of the L1 site early in infection. MLTU site selection may proceed in a first-come, first-serve manner whereby the L1 site is used most frequently because it is closest to the promoter. Alternatively, specific sequences flanking the L1 site may control predominant L1 site usage in a position-independent manner. To distinguish between these mechanisms, we constructed deletions in the L1 flanking sequences and inserted the mutated sites into either simple transcription units or mini-MLTUs encoding two poly(A) sites. The pattern of site selection for each construct was then quantitated by S1 nuclease analysis after transfection into 293 cells. The results indicated that L1 sequences upstream of AAUAAA define a novel selector element that can cause predominant L1 site usage at either position of a tandem transcription unit. The element did not significantly affect the stability or nucleocytoplasmic transport of L1 transcripts and was not required for efficient 3'-end processing in simple transcription units. Predominant L1 site usage required physical linkage of the processing signals and was independent of the major late promoter.
AbstractList The adenovirus major late transcription unit (MLTU) encodes five colinear mRNA families, L1 through L5, each distinguished by a unique poly(A) site. Site selection is regulated during the course of infection, predominating early at the L1 site and late at the L2 through L5 sites. Two general mechanisms can be invoked to explain predominant usage of the L1 site early in infection. MLTU site selection may proceed in a first-come, first-serve manner whereby the L1 site is used most frequently because it is closest to the promoter. Alternatively, specific sequences flanking the L1 site may control predominant L1 site usage in a position-independent manner. To distinguish between these mechanisms, we constructed deletions in the L1 flanking sequences and inserted the mutated sites into either simple transcription units or mini-MLTUs encoding two poly(A) sites. The pattern of site selection for each construct was then quantitated by S1 nuclease analysis after transfection into 293 cells. The results indicated that L1 sequences upstream of AAUAAA define a novel selector element that can cause predominant L1 site usage at either position of a tandem transcription unit. The element did not significantly affect the stability or nucleocytoplasmic transport of L1 transcripts and was not required for efficient 3'-end processing in simple transcription units. Predominant L1 site usage required physical linkage of the processing signals and was independent of the major late promoter.
Author Imperiale, M J
DeZazzo, J D
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Snippet The adenovirus major late transcription unit (MLTU) encodes five colinear mRNA families, L1 through L5, each distinguished by a unique poly(A) site. Site...
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StartPage 4951
SubjectTerms Adenoviridae - genetics
Base Sequence
Biological Transport
Cell Line
DNA Mutational Analysis
Genetic Linkage
Humans
Plasmids
Poly A - metabolism
Promoter Regions, Genetic
Restriction Mapping
RNA, Messenger
Transcription, Genetic
Title Sequences upstream of AAUAAA influence poly(A) site selection in a complex transcription unit
URI https://www.ncbi.nlm.nih.gov/pubmed/2601703
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