Molecular and biochemical analysis of the α class carbonic anhydrases in Caenorhabditis elegans

Abstract

In this study, in silico analysis of the Caenorhabditis elegans genome revealed six genes (cah-1, cah-2, cah-3, cah-4, cah-5, and cah-6) possibly encoding α class CAs (carbonic anhydrase). Real-time RT-PCR analysis revealed the temporal expression pattern of each gene, as well as changes in expression levels under different atmospheric conditions (stress). Cah-3 and cah-4 showed the highest levels of transcript accumulation, while most genes responded to the stress conditions. Yeast complementation showed that cah-3 was able to complement the function of Saccharomyces cerevisiae CA (NCE103) in vivo. Recombinant CAH-3, CAH-4a and CAH-5 enzymes, expressed in Escherichia coli were used for in vitro measurement of CA activity. However, in vitro activity was only detectable for CAH-4a. RNAi by feeding was performed on wild-type C. elegans for all genes. The worms were examined for a visible phenotype under normal and stress conditions (pH, CO(2)/O(2)). Silencing cah-3 and cah-4 may reduce the life-span of the worms (at 22 °C).

CAS-1, a C. elegans cyclase-associated protein, is required for sarcomeric actin assembly in striated muscle

“…Later, this gene was renamed as cas-1 (cyclase-associated protein-1) because the gene name 'cah' was suited better to represent the carbonic anhydrase gene family. As a result, currently known cah-1 is a gene that encodes a carbonic anhydrase (Fasseas et al, 2011) and unrelated to cas-1, which is the major subject of the current study. Although 'CAP' is the common name for this protein family, the gene name 'cap' has already been used for designation of C. elegans genes encoding heterodimeric actin capping protein subunits (cap-1 and cap-2) (Waddle et al, 1993).…”

Drosophila melanogaster: a model organism for controllingDipteranvectors and pests

“…In protozoan and metazoan, only few studies have been conducted on b-CAs. The majority of studies are related to cloning, characterization, and functional analysis of a-CAs, including Caenorhabditis elegans 43 , Ostertagia ostertagi (a nematode and causative agent of Ostertagiosis in goat, sheep, cow, and cattle) 44 , Plasmodium falciparum (causative agent of malaria) 45 , Riftia pachyptila (giant tube worm) 46 , and Trypanosoma cruzi (causative agent of Chagas disease) 47 . In 2010, two independent studies initiated investigations of b-CAs in protozoans and metazoans.…”

“…In 2010, two independent studies initiated investigations of b-CAs in protozoans and metazoans. Two genes encoding b-CAs (y116a8c.28 and bca-1) were identified in Caenorhabditis elegans 48 . Our group reported a novel b-CA gene identified in FlyBase (http://flybase.org/), which was named D. melanogaster (fruit fly) b-CA (DmBCA).…”

Identification of a nuclear carbonic anhydrase in Caenorhabditis elegans

“…Thus, C. elegans may be a useful reductionist model for discerning the function of both catalytic CAs as well as the evolutionarily conserved acatalytic proteins. Previously, the temporal expression patterns of the worm CA gene family has been studied [29] and one of these genes, cah-4 , has been shown to be regulated by both environmental pH [30] and oxygen levels [31]. Cah-4 may also play a role in the progression of muscle degeneration in a worm model of muscular dystrophy [32].…”