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Mitochondrial ribosome assembly in Neurospora. Structural analysis of mature and partially assembled ribosomal subunits by equilibrium centrifugation in CsCl gradients

LaPolla, Robert J. and Lambowitz, Alan M. (1982) Mitochondrial ribosome assembly in Neurospora. Structural analysis of mature and partially assembled ribosomal subunits by equilibrium centrifugation in CsCl gradients. Journal of Cell Biology, 95 (1). pp. 267-277. ISSN 0021-9525. http://resolver.caltech.edu/CaltechAUTHORS:LAPjcb82

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Abstract

In Neurospora, one protein associated with the mitochondrial small ribosomal subunit (S-5, Mr 52,000) is synthesized intramitochondrially and is assumed to be encoded by mtDNA. When mitochondrial protein synthesis is inhibited, either by chloramphenicol or by mutation, cells accumulate incomplete mitochondrial small subunits (CAP-30S and INC-30S particles) that are deficient in S-5 and several other proteins. To gain additional insight into the role of S-5 in mitochondrial ribosome assembly, the structures of Neurospora mitochondrial ribosomal subunits, CAP-30S particles, and INC-30S particles were analyzed by equilibrium centrifugation in CsCl gradients containing different concentrations of Mg+2. The results show (a) that S-5 is tightly associated with small ribosomal subunits, as judged by the fact that it is among the last proteins to be dissociated in CsCl gradients as the Mg+2 concentration is decreased, and (b) that CAP-30S and INC-30S particles, which are deficient in S-5, contain at most 12 proteins that are bound as tightly as in mature small subunits. The CAP-30S particles isolated from sucrose gradients contain a number of proteins that appear to be loosely bound, as judged by dissociation of these proteins in CsCl gradients under conditions in which they remain associated with mature small subunits. The results suggest that S-5 is required for the stable binding of a subset of small subunit ribosomal proteins.


Item Type:Article
Additional Information:Copyright © 1982 by The Rockefeller University Press. Received for publication 9 April 1982, and in revised form 2 July 1982. We thank Mr. Kenneth Figlioli for technical assistance. This work was supported by National Institutes of Health grant GM 23961.
Record Number:CaltechAUTHORS:LAPjcb82
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:LAPjcb82
Alternative URL:http://www.jcb.org/cgi/content/abstract/95/1/267
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:9980
Collection:CaltechAUTHORS
Deposited By: Archive Administrator
Deposited On:01 Apr 2008
Last Modified:26 Dec 2012 09:55

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