Biochemical and molecular characterization of two cytidine deaminases in the nematode Caenorhabditis elegans

Two cytidine deaminases (CDDs) from the free-living nematode Caenorhabditis elegans have been cloned and characterized. Both Ce-CDD-1 and Ce-CDD-2 are authentic deaminases and both exhibit RNA-binding activity towards AU-rich templates. In order to study their temporal and spatial expression pattern...

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Bibliographic Details
Published in:Biochemical journal 2002-07, Vol.365 (Pt 1), p.99-107
Main Authors: Thompson, Fiona J, Britton, Collette, Wheatley, Isla, Maitland, Kirsty, Walker, Glenda, Anant, Shrikant, Davidson, Nicholas O, Devaney, Eileen
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Caenorhabditis elegans - enzymology
Caenorhabditis elegans - genetics
Cloning, Molecular
Cytidine Deaminase - antagonists & inhibitors
Cytidine Deaminase - chemistry
Cytidine Deaminase - genetics
Cytidine Deaminase - metabolism
DNA, Helminth - genetics
Female
Gene Expression Regulation, Developmental
Gene Expression Regulation, Enzymologic
Genes, Helminth
Genes, Reporter
Intestines - enzymology
Isoenzymes - antagonists & inhibitors
Isoenzymes - chemistry
Isoenzymes - genetics
Isoenzymes - metabolism
Molecular Sequence Data
Oogenesis - genetics
Oogenesis - physiology
Phenotype
Sequence Homology, Amino Acid
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Summary:Two cytidine deaminases (CDDs) from the free-living nematode Caenorhabditis elegans have been cloned and characterized. Both Ce-CDD-1 and Ce-CDD-2 are authentic deaminases and both exhibit RNA-binding activity towards AU-rich templates. In order to study their temporal and spatial expression patterns in the worm, reporter gene constructs were made using approx. 2 kb of upstream sequence. Transfection of C. elegans revealed that both genes localized to the cells of the intestine, although their temporal expression patterns were different. Expression of Ce-cdd-1 peaked in the early larval stages, whereas Ce-cdd-2 was expressed in all life cycle stages examined. RNA-interference (RNAi) assays were performed for both genes, either alone or in combination, but only cdd-2 RNAi produced a consistent visible phenotype. A proportion of eggs laid from these worms were swollen and distorted in shape.
ISSN:0264-6021
1470-8728
DOI:10.1042/BJ20011814