Histone-specific RNA 3' processing in nuclear extracts from mammalian cells.

Stauber, C; Soldati, D; Lüscher, B; Schümperli, Daniel (1990). Histone-specific RNA 3' processing in nuclear extracts from mammalian cells. In: RNA Processing Part B: Specific Methods. Methods in enzymology: Vol. 181 (pp. 74-89). San Diego, CA: Academic Press

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The mature 3' ends of histone mRNAs are formed by endonucleolytic cleavage of longer precursor transcripts. This process occurs in the nucleus and can be regarded as the equivalent of the polyadenylation reaction involved in 3′-end-generation of all other mRNAs. A sea urchin H3 gene that failed to be properly processed in the Xenopus oocyte system proved particularly useful, because it allowed the identification of a processing component from sea urchins by a complementation assay. Nuclear extracts prepared from cells under various growth conditions have helped to reveal proliferation-dependent changes in the efficiency of histone RNA 3′ processing. RNA substrates for in vitro processing are best prepared by runoff transcription of specific DNA templates with bacterial or phage RNA polymerases. For this purpose, a restriction fragment containing the 3′-terminal region of a histone gene and including the conserved palindrome and spacer motifs is cloned into a polylinker sequence downstream of a strong promoter.

Item Type:	Book Section (Book Chapter)
Division/Institute:	08 Faculty of Science > Department of Biology > Institute of Cell Biology 08 Faculty of Science > Department of Biology > Institute of Cell Biology > RNA
UniBE Contributor:	Schümperli, Daniel
Subjects:	500 Science > 570 Life sciences; biology
ISSN:	0076-6879
ISBN:	978-0-12-182082-4
Series:	Methods in enzymology
Publisher:	Academic Press
Language:	English
Submitter:	Daniel Schümperli
Date Deposited:	10 Mar 2016 15:18
Last Modified:	05 Dec 2022 14:52
PubMed ID:	2166220
URI:	https://boris.unibe.ch/id/eprint/77097