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Evidence for translational control of β-tubulin synthesis during differentiation of Leishmania donovani

Published online by Cambridge University Press:  06 April 2009

M. Bhaumik ,
S. Das  and
S. Adhya
M. Bhaumik
Affiliation:
Genetic Engineering Laboratory, Leishmania Group, Indian Institute of Chemical Biology, 4, Raja S.C. Mullick Road, Calcutta-700 032, India
S. Das
Affiliation:
Genetic Engineering Laboratory, Leishmania Group, Indian Institute of Chemical Biology, 4, Raja S.C. Mullick Road, Calcutta-700 032, India
S. Adhya
Affiliation:
Genetic Engineering Laboratory, Leishmania Group, Indian Institute of Chemical Biology, 4, Raja S.C. Mullick Road, Calcutta-700 032, India
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Abstract

Tubulin biosynthesis was rapidly induced during transformation of the mammalian (amastigote) stage of the kinetoplastid parasite Leishmania donovani to flagellated promastigotes. However, transcription of β-tubulin genes occurred constitutively, as judged by nascent RNA synthesis in isolated nuclei and Northern blotting of steady-state mRNA. Two mRNA species of 2.2 and 2.4 kb were shared by the two cell-types, while a third 2.6 kb species, constituting about 20% of the total, was present in large amounts in promastigotes. RNase protection experiments demonstrated sequence micro-heterogeneity in the 5′-untranslated region, the pattern of which was identical in promastigotes and amastigotes. By primer extension assays, heterogeneity in the 5′-terminal cap structure of amastigote β-tubulin mRNA and differential pausing of reverse transcriptase within the mini-exon leader region were detected. These differences correlated with enhanced translational efficiency of tubulin mRNA from promastigotes in a rabbit reticulocyte lysate system. The results indicate that translational control plays a major role in tubulin induction during L. donovani differentiation.

Keywords

β-tubulin mRNAtransformationLeishmania donovani
Type
Research Article
Information
Parasitology , Volume 103 , Issue 2 , October 1991 , pp. 197 - 205
Copyright
Copyright © Cambridge University Press 1991

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