Cotranscriptional processing of Drosophila histone mRNAs

Todd E Adamson; David H Price

doi:10.1128/MCB.23.12.4046-4055.2003

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Cotranscriptional processing of Drosophila histone mRNAs

Journal article

Peer reviewed

Cotranscriptional processing of Drosophila histone mRNAs

Todd E Adamson and David H Price

Molecular and cellular biology, Vol.23(12), pp.4046-4055

06/2003

DOI: 10.1128/MCB.23.12.4046-4055.2003

PMCID: PMC156150

PMID: 12773550

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http://doi.org/10.1128/MCB.23.12.4046-4055.2003View

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Abstract

The 3' ends of metazoan histone mRNAs are generated by specialized processing machinery that cleaves downstream of a conserved stem-loop structure. To examine how this reaction might be influenced by transcription, we used a Drosophila melanogaster in vitro system that supports both processes. In this system the complete synthesis of histone mRNA, including transcription initiation and elongation, followed by 3' end formation, occurred at a physiologically significant rate. Processing of free transcripts was efficient and occurred with a t(1/2) of less than 1 min. Divalent cations were not required, but nucleoside triphosphates (NTPs) stimulated the rate of cleavage slightly. Isolated elongation complexes encountered a strong arrest site downstream of the mature histone H4 3' end. In the presence of NTPs, transcripts in these arrested complexes were processed at a rate similar to that of free RNA. Removal of NTPs dramatically reduced this rate, potentially due to concealment of the U7 snRNP binding element. The arrest site was found to be a conserved feature located 32 to 35 nucleotides downstream of the processing site on the H4, H2b, and H3 genes. The significance of the newly discovered arrest sites to our understanding of the coupling between transcription and RNA processing on the one hand and histone gene expression on the other is discussed.

Biochemistry

Polymerase Chain Reaction

Cell Line

Gene Expression Regulation

DNA - metabolism

RNA, Messenger - metabolism

Drosophila melanogaster - metabolism

Animals

Cell Nucleus - metabolism

Time Factors

Models, Biological

Transcription, Genetic

Biochemical Phenomena

Models, Genetic

Histones - metabolism

Kinetics

RNA Polymerase II - chemistry

RNA - metabolism

Details

Title: Subtitle: Cotranscriptional processing of Drosophila histone mRNAs
Creators: Todd E Adamson - Department of Biochemistry, University of Iowa, Iowa City, Iowa 52242, USA
David H Price
Resource Type: Journal article
Publication Details: Molecular and cellular biology, Vol.23(12), pp.4046-4055
DOI: 10.1128/MCB.23.12.4046-4055.2003
PMID: 12773550
PMCID: PMC156150
NLM abbreviation: Mol Cell Biol
ISSN: 0270-7306
eISSN: 1098-5549
Publisher: United States
Grant note: GM35500 / NIGMS NIH HHS R01 GM035500 / NIGMS NIH HHS
Language: English
Date published: 06/2003
Academic Unit: Biochemistry and Molecular Biology
Record Identifier: 9984024554702771

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