Cloning and expression analysis of Drosophila extracellular Cu Zn superoxide dismutase
Cloning and expression analysis of Drosophila extracellular Cu Zn superoxide dismutase
In the present study, we cloned and sequenced the mRNAs of the Sod3 [extracellular Cu Zn SOD (superoxide dismutase)] gene in Drosophila and identified two mRNA products formed by alternative splicing. These products code for a long and short protein derived from the four transcripts found in global expression studies (Flybase numbers Dmel\CG9027, FBgn0033631). Both mRNA process variants contain an extracellular signalling sequence, a region of high homology to the Sod1 (cytoplasmic Cu Zn SOD) including a conserved AUG start, with the longer form also containing a hydrophobic tail. The two fully processed transcripts are homologous to Caenorhabditis elegans Sod3 mRNA showing the same processing pattern. Using an established KG p-element+ insertion line (KG06029), we demonstrate that the Sod3 codes for an active Cu Zn SOD. We found differing expression patterns across sex with higher levels of expression of Sod3 in females. There is a correlation of Sod1 and Sod3 gene expression and activity that can explain why Sod3 was not seen in earlier studies of Sod1. Finally, we found no effect on lifespan with the Sod3 hypomorph mutation (Sod3KG06029) but did observe a significant increase in resistance to paraquat and H2O2 (hydrogen peroxide).
Amino Acid Sequence, Animals, Cloning, Molecular, Drosophila Proteins/genetics, Drosophila melanogaster/drug effects, Extracellular Space/enzymology, Female, Gene Expression Regulation, Enzymologic, Herbicides/pharmacology, Hydrogen Peroxide/pharmacology, Isoenzymes/genetics, Male, Molecular Sequence Data, Mutation, Oxidants/pharmacology, Paraquat/pharmacology, Reverse Transcriptase Polymerase Chain Reaction, Sequence Homology, Amino Acid, Sex Factors, Superoxide Dismutase/genetics, Survival Analysis
Blackney, Michael J
6aa91b37-4f60-4858-a5cf-fde0bfedfa66
Cox, Rebecca
c0677628-088b-4af1-844e-da7c78ab72eb
Shepherd, David
11aa6858-d19c-4450-82ff-11dff9dcd9c4
Parker, Joel D
23c6e137-cc5c-4ea1-a9b2-c87d750a68f5
23 December 2014
Blackney, Michael J
6aa91b37-4f60-4858-a5cf-fde0bfedfa66
Cox, Rebecca
c0677628-088b-4af1-844e-da7c78ab72eb
Shepherd, David
11aa6858-d19c-4450-82ff-11dff9dcd9c4
Parker, Joel D
23c6e137-cc5c-4ea1-a9b2-c87d750a68f5
Blackney, Michael J, Cox, Rebecca, Shepherd, David and Parker, Joel D
(2014)
Cloning and expression analysis of Drosophila extracellular Cu Zn superoxide dismutase.
Bioscience Reports, 34 (6), [e00164].
(doi:10.1042/BSR20140133).
Abstract
In the present study, we cloned and sequenced the mRNAs of the Sod3 [extracellular Cu Zn SOD (superoxide dismutase)] gene in Drosophila and identified two mRNA products formed by alternative splicing. These products code for a long and short protein derived from the four transcripts found in global expression studies (Flybase numbers Dmel\CG9027, FBgn0033631). Both mRNA process variants contain an extracellular signalling sequence, a region of high homology to the Sod1 (cytoplasmic Cu Zn SOD) including a conserved AUG start, with the longer form also containing a hydrophobic tail. The two fully processed transcripts are homologous to Caenorhabditis elegans Sod3 mRNA showing the same processing pattern. Using an established KG p-element+ insertion line (KG06029), we demonstrate that the Sod3 codes for an active Cu Zn SOD. We found differing expression patterns across sex with higher levels of expression of Sod3 in females. There is a correlation of Sod1 and Sod3 gene expression and activity that can explain why Sod3 was not seen in earlier studies of Sod1. Finally, we found no effect on lifespan with the Sod3 hypomorph mutation (Sod3KG06029) but did observe a significant increase in resistance to paraquat and H2O2 (hydrogen peroxide).
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bsr034e164
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Published date: 23 December 2014
Keywords:
Amino Acid Sequence, Animals, Cloning, Molecular, Drosophila Proteins/genetics, Drosophila melanogaster/drug effects, Extracellular Space/enzymology, Female, Gene Expression Regulation, Enzymologic, Herbicides/pharmacology, Hydrogen Peroxide/pharmacology, Isoenzymes/genetics, Male, Molecular Sequence Data, Mutation, Oxidants/pharmacology, Paraquat/pharmacology, Reverse Transcriptase Polymerase Chain Reaction, Sequence Homology, Amino Acid, Sex Factors, Superoxide Dismutase/genetics, Survival Analysis
Identifiers
Local EPrints ID: 455407
URI: http://eprints.soton.ac.uk/id/eprint/455407
ISSN: 0144-8463
PURE UUID: 6f4299da-b040-4c70-a175-750fdb5b3540
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Date deposited: 21 Mar 2022 17:36
Last modified: 06 Aug 2024 01:52
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Author:
Michael J Blackney
Author:
Rebecca Cox
Author:
Joel D Parker
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