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PMID: 976015 Published · ppublish English Journal Article

In vitro transcription by isolated nuclei of Rhynchosciara americana salivary glands. Characteristics of incorporation and inhibition by alpha-amanitin.

Chromosoma ·Vol. 56 ·No. 1 ·1976-06-30 ·Pages 69-84

Santelli RV, Machado-Santelli GM, Lara FJ

Abstract

A method for the isolation of polytene nuclei from salivary glands cells of the Diptera Rhynchosciara americana is described. The stage-specific morphological pattern of the chromosome is maintained during the isolation. The isolated nuclei show two distinct RNA polymerase activities, namely I and II, characterized on the basis of ionic requirements and alpha-amanitin sensitivity. Studies of the product under the incubation conditions show that the system allows the synthesis of high-molecular weight RNA, beside a low molecular weight peak which may comprise pre-4S and 5S RNAs. Autoradiographic studies carried out in the presence or absence of the toxin alpha-amanitin showed that micronucleoli contain products of RNA polymerase type I activity (ribosomal RNA) and that the DNA puffs are engaged in alpha-amanitin sensitive RNA synthesis and thus are sites of polymerase type II activity.

MeSH Terms
Amanitins/pharmacology Animals Autoradiography Cell Fractionation/methods Cell Nucleolus/enzymology Cell Nucleus/metabolism DNA-Directed RNA Polymerases/analysis,antagonists & inhibitors Diptera/cytology Osmolar Concentration RNA/biosynthesis RNA, Ribosomal/biosynthesis Salivary Glands/ultrastructure Transcription, Genetic/drug effects
Chemicals
Amanitins RNA, Ribosomal RNA DNA-Directed RNA Polymerases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Santelli R V
Machado-Santelli G M
Lara F J
References (43)
43 references, click to expand
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Article Info
Journal
Chromosoma
Abbr.
Chromosoma
ISSN
0009-5915
DOI
10.1007/BF00293731
Published
1976-06-30
Pages
69-84
Language
English
Region
Austria
NLM ID
2985138R
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00293731 View journal details
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