TY - JOUR

T1 - Cloning and expression analysis of Drosophila extracellular Cu Zn superoxide dismutase

AU - Blackney, Michael J.

AU - Cox, Rebecca

AU - Shepherd, David

AU - Parker, Joel D.

PY - 2014/12/23

Y1 - 2014/12/23

N2 - 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).

AB - 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).

KW - Amino Acid Sequence

KW - Animals

KW - Cloning, Molecular

KW - Drosophila Proteins/genetics

KW - Drosophila melanogaster/drug effects

KW - Extracellular Space/enzymology

KW - Female

KW - Gene Expression Regulation, Enzymologic

KW - Herbicides/pharmacology

KW - Hydrogen Peroxide/pharmacology

KW - Isoenzymes/genetics

KW - Male

KW - Molecular Sequence Data

KW - Mutation

KW - Oxidants/pharmacology

KW - Paraquat/pharmacology

KW - Reverse Transcriptase Polymerase Chain Reaction

KW - Sequence Homology, Amino Acid

KW - Sex Factors

KW - Superoxide Dismutase/genetics

KW - Survival Analysis

U2 - 10.1042/BSR20140133

DO - 10.1042/BSR20140133

M3 - Article

C2 - 25339624

VL - 34

SP - e00164

JO - Bioscience reports

JF - Bioscience reports

SN - 0144-8463

IS - 6

ER -