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Pleiotropic consequences of misexpression of the developmentally active and stress-inducible non-coding hsrω gene in Drosophila

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Abstract

The non-coding hsrω gene of Drosophila melanogaster is expressed in nearly all cell types and developmental stages. However, in the absence of conventional mutant alleles of this gene, its developmental functions remain largely unknown. In the present study, we used a variety of GAL4 drivers to overexpress or ablate this gene’s transcripts in specific tissues and examined the developmental consequences thereof. Our results show that a balanced expression of these non-coding transcripts is critical for survival and normal development in all the tissue types tested, since any change in cellular levels of these transcripts in a given cell type generally has detrimental effects, with extreme cases resulting in organismal lethality, although in a few cases the misexpression of these transcripts also suppresses the mutant phenotype due to other genetic conditions. Evidence is also presented for existence of a new spliced variant of the hsrω-n nuclear transcript. Following the RNAi-mediated down-regulation of hsrω transcripts, the omega speckles disappear so that the nucleoplasmic hnRNPs get diffusely distributed, while up-regulation of these transcripts results in greater sequestration of these proteins into omega speckle clusters; either of these conditions would affect activities of the hnRNPs and other hsrω-RNA interacting proteins, which is likely to have cascading consequences. The present findings, together with our earlier observations on effects of altered levels of the hsrω transcripts on induced apoptosis and expanded polyQ-mediated neurodegeneration, further confirm that ncRNA species like the hsrω, far from being evolutionary hangovers, provide critical information for important functions in normal cells.

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Abbreviations

AP or A/P:

anterior-posterior

DIG:

digoxigenin

EP:

enhancer promoter

FRISH:

fluorescent RNA:RNA in situ hybridization

hnRNP:

heterogeneous nuclear RNA-binding protein

HP1:

heterochromatin protein 1

IGC:

inter-chromatin granule cluster

ncRNA:

non-coding RNA

ORF:

open reading frame

polyQ:

polyglutamine

RISC:

RNA-induced silencing complex

RNAi:

RNA interference

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Acknowledgements

We are thankful to the Bloomington Stock Center for EP3037, dpp-GAL4, ptc-GAL4, P{GawB}ap md544 , P{GawB}l(2)C805 C805, P{w[+mW.hs] = GawB}c825, hs-GAL4 t , Act5C-GAL4, elav-GAL4 and GMR-GAL4 fly stocks and to Dr VK Parnaik for providing the UAS-lamC flies. We gratefully thank Dr H Saumweber for generously providing the P11 antibody. This work was supported, in part, by the Shyama Prasad Mukherjee Fellowship from the Council of Scientific and Industrial Research, India, to MM and research support from the Department of Science and Technology, India, to SCL. The confocal microscope facility is also funded by the Department of Science and Technology, India.

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  1. Moushami Mallik

    Present address: Present address: Molecular Neurogenetics Laboratory, Max-Planck Institute for Molecular Biomedicine, Roentgenstrasse 20, 48149, Muenster, Germany

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  1. Cytogenetics Laboratory, Department of Zoology, Banaras Hindu University, Varanasi, 221 005, India

    Moushami Mallik & Subhash C Lakhotia

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  1. Moushami Mallik
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Correspondence to Subhash C Lakhotia.

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Mallik, M., Lakhotia, S.C. Pleiotropic consequences of misexpression of the developmentally active and stress-inducible non-coding hsrω gene in Drosophila . J Biosci 36, 265–280 (2011). https://doi.org/10.1007/s12038-011-9061-x

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