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The ribosomes of Plasmodium berghei: isolation and ribosomal ribonucleic acid analysis

Published online by Cambridge University Press:  06 April 2009

F. W. Miller  and
Judith Ilan
F. W. Miller
Affiliation:
Department of Anatomy and Developmental Biology Center, Case Western Reserve University, Cleveland, Ohio 44106
Judith Ilan*
Affiliation:
Department of Anatomy and Developmental Biology Center, Case Western Reserve University, Cleveland, Ohio 44106
*
Dr Judith Ilan, Department of Anatomy, School of Medicine, Case Western Reserve University, Cleveland, Oh 44106
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Summary

Ribosomes and high molecular weight ribosomal ribonucleic acid (rRNA) from the blood stages of Plasmodium berghei parasites were studied in preparations free from host ribosome contamination. Purified malarial ribosomes were isolated in high yield from a population of ultrastructurally intact, viable parasites by hypertonic lysis with Triton X-100 and differential centrifugation. These ribosomes were shown to be derived from active polysomes and could be dissociated into subunits by puromycin–0·5 m KCl treatment. Malarial rRNA extracted from purified 40S and 60S ribosomal subunits was characterized by electrophoretic, sedimentation and base ratio analyses. Like certain other protozoa, the P. berghei 40S ribosomal subunit possessed an exceptionally large RNA species (mol. wt 0·9 × 106), while RNA isolated from the parasite's 60S subunit (mol. wt 1·5 × 106) was specifically ‘nicked’ to produce one large component (mol.wt 1·2 × 106) and one small component (mol.wt 0·3 × 106) in equimolar quantities. These rRNA's migrate identically on polyacrylamide gels after heating to 63°C for 5 mm or under denaturing conditions in the presence of formamide, indicating an absence of aggregation and non-specific degradation of the rRNA species. Base composition studies showed P. berghei rRNA to be low in guanosine and cytosine content, as is the case for protozoa generally.

Type
Research Article
Information
Parasitology , Volume 77 , Issue 3 , December 1978 , pp. 345 - 366
Copyright
Copyright © Cambridge University Press 1978

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