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Subdued, a TMEM16 family Ca²⁺-activated Cl⁻channel in Drosophila melanogaster with an unexpected role in host defense.

Wong XM, Younger S, Peters CJ, Jan YN, Jan LY - Elife (2013)

Bottom Line: Whether distant relatives of the vertebrate TMEM16 families also form CaCCs is an intriguing open question.Amino acid substitutions of Subdued alter the ion selectivity and kinetic properties of the CaCC channels heterologously expressed in HEK 293T cells.This Drosophila channel displays characteristics of classic CaCCs, thereby providing evidence for evolutionarily conserved biophysical properties in the TMEM16 family.

View Article: PubMed Central - PubMed

Affiliation: Graduate Program in Chemistry and Chemical Biology, University of California, San Francisco, San Francisco, United States.

ABSTRACT
TMEM16A and TMEM16B are calcium-activated chloride channels (CaCCs) with important functions in mammalian physiology. Whether distant relatives of the vertebrate TMEM16 families also form CaCCs is an intriguing open question. Here we report that a TMEM16 family member from Drosophila melanogaster, Subdued (CG16718), is a CaCC. Amino acid substitutions of Subdued alter the ion selectivity and kinetic properties of the CaCC channels heterologously expressed in HEK 293T cells. This Drosophila channel displays characteristics of classic CaCCs, thereby providing evidence for evolutionarily conserved biophysical properties in the TMEM16 family. Additionally, we show that knockout flies lacking subdued gene activity more readily succumb to death caused by ingesting the pathogenic bacteria Serratia marcescens, suggesting that subdued has novel functions in Drosophila host defense. DOI: http://dx.doi.org/10.7554/eLife.00862.001.

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CG16718 (Subdued) aligns closely with mammalian CaCCs TMEM16A and B (Mus musculus).(A) Multiple sequence alignment of protein sequences was done with ClustalW2 and phylogenetic tree construction was done in PHYLIP 3.67 (Drawgram). (B) Putative transmembrane segments are highlighted with boxes, and mutated residues (Figure 4) featured in this report are marked in color on the primary sequence alignment Y489 is shown in green and Q672 is shown in red. Bracketed values at the end of the alignment indicate the number of residues in the whole channel.DOI:http://dx.doi.org/10.7554/eLife.00862.003
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fig1: CG16718 (Subdued) aligns closely with mammalian CaCCs TMEM16A and B (Mus musculus).(A) Multiple sequence alignment of protein sequences was done with ClustalW2 and phylogenetic tree construction was done in PHYLIP 3.67 (Drawgram). (B) Putative transmembrane segments are highlighted with boxes, and mutated residues (Figure 4) featured in this report are marked in color on the primary sequence alignment Y489 is shown in green and Q672 is shown in red. Bracketed values at the end of the alignment indicate the number of residues in the whole channel.DOI:http://dx.doi.org/10.7554/eLife.00862.003

Mentions: Multiple sequence alignment with the mammalian TMEM16 family shows that Subdued is most similar to TMEM16A and B (Figure 1A,B), sharing 32.8% and 31.7% identity with these channels, respectively.10.7554/eLife.00862.003Figure 1.CG16718 (Subdued) aligns closely with mammalian CaCCs TMEM16A and B (Mus musculus).


Subdued, a TMEM16 family Ca²⁺-activated Cl⁻channel in Drosophila melanogaster with an unexpected role in host defense.

Wong XM, Younger S, Peters CJ, Jan YN, Jan LY - Elife (2013)

CG16718 (Subdued) aligns closely with mammalian CaCCs TMEM16A and B (Mus musculus).(A) Multiple sequence alignment of protein sequences was done with ClustalW2 and phylogenetic tree construction was done in PHYLIP 3.67 (Drawgram). (B) Putative transmembrane segments are highlighted with boxes, and mutated residues (Figure 4) featured in this report are marked in color on the primary sequence alignment Y489 is shown in green and Q672 is shown in red. Bracketed values at the end of the alignment indicate the number of residues in the whole channel.DOI:http://dx.doi.org/10.7554/eLife.00862.003
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3814000&req=5

fig1: CG16718 (Subdued) aligns closely with mammalian CaCCs TMEM16A and B (Mus musculus).(A) Multiple sequence alignment of protein sequences was done with ClustalW2 and phylogenetic tree construction was done in PHYLIP 3.67 (Drawgram). (B) Putative transmembrane segments are highlighted with boxes, and mutated residues (Figure 4) featured in this report are marked in color on the primary sequence alignment Y489 is shown in green and Q672 is shown in red. Bracketed values at the end of the alignment indicate the number of residues in the whole channel.DOI:http://dx.doi.org/10.7554/eLife.00862.003
Mentions: Multiple sequence alignment with the mammalian TMEM16 family shows that Subdued is most similar to TMEM16A and B (Figure 1A,B), sharing 32.8% and 31.7% identity with these channels, respectively.10.7554/eLife.00862.003Figure 1.CG16718 (Subdued) aligns closely with mammalian CaCCs TMEM16A and B (Mus musculus).

Bottom Line: Whether distant relatives of the vertebrate TMEM16 families also form CaCCs is an intriguing open question.Amino acid substitutions of Subdued alter the ion selectivity and kinetic properties of the CaCC channels heterologously expressed in HEK 293T cells.This Drosophila channel displays characteristics of classic CaCCs, thereby providing evidence for evolutionarily conserved biophysical properties in the TMEM16 family.

View Article: PubMed Central - PubMed

Affiliation: Graduate Program in Chemistry and Chemical Biology, University of California, San Francisco, San Francisco, United States.

ABSTRACT
TMEM16A and TMEM16B are calcium-activated chloride channels (CaCCs) with important functions in mammalian physiology. Whether distant relatives of the vertebrate TMEM16 families also form CaCCs is an intriguing open question. Here we report that a TMEM16 family member from Drosophila melanogaster, Subdued (CG16718), is a CaCC. Amino acid substitutions of Subdued alter the ion selectivity and kinetic properties of the CaCC channels heterologously expressed in HEK 293T cells. This Drosophila channel displays characteristics of classic CaCCs, thereby providing evidence for evolutionarily conserved biophysical properties in the TMEM16 family. Additionally, we show that knockout flies lacking subdued gene activity more readily succumb to death caused by ingesting the pathogenic bacteria Serratia marcescens, suggesting that subdued has novel functions in Drosophila host defense. DOI: http://dx.doi.org/10.7554/eLife.00862.001.

Show MeSH
Related in: MedlinePlus