TY - JOUR
T1 - CHCA-1 is a copper-regulated CTR1 homolog required for normal development, copper accumulation, and copper-sensing behavior in Caenorhabditis elegans
AU - Yuan, Sai
AU - Sharma, Anuj Kumar
AU - Richart, Alexandria
AU - Lee, Jaekwon
AU - Kim, Byung Eun
N1 - Funding Information:
This work was supported in part by National Institutes of Health Grants DK110195 (to B.-E. K.) and DK079209 (to J. L.) and Nebraska Redox Biology Center Grant P30GM103335 (to J. L.). The authors declare that they have no conflicts of interest with the contents of this article. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Funding Information:
C. elegans were cultured at 20 °C on NGM plates seeded with Escherichia coli OP50 for general maintenance or with E. coli HT115 dsRNA-expressing bacteria for RNAi experiments (58). Bristol N2 was used as the WT C. elegans strain. Mutant and transgenic strains were outcrossed with N2 to obtain WT backgrounds, and a WT brood mate animal was used following crossing in mutant and transgenic animal growth/avoidance assays. Some strains were provided by the CGC, which is funded by National Institutes of Health Office of Research Infrastructure Programs (P40 OD010440). The chca-1 (tm6506) IV strain was obtained from the National Bioresource Project (33) and outcrossed with N2 six times prior to use to establish heritability; CB1033 (che-2 (e1033) X) was obtained from CGC and outcrossed six times before use for the same purpose. A list of transgenic worms used in this study can be found in Table S2. Transgenic animals in a chca-1 (tm6506) mutant background, as well as multiple transgene-presenting strains, were generated with standard mating methods; genotypes were confirmed by PCR and/or DNA sequencing. The chca-1 (tm6506) genotyping primers were 5′-GTATCTAGTCCGATAAGAAG-3′ and 5′-TTGAAGCAAAAACAAAGTGC-3′.
Funding Information:
Acknowledgments—We thank members of the Kim laboratory for helpful suggestions, technical assistance, and critical reading of this manuscript. We thank members of Dr. Iqbal Hamza’s laboratory (University of Maryland, College Park) for technical assistance and expertise in C. elegans genetics and cell biology as well as for discussions regarding the work, and Dr. Tamara Korolnek for critical reading of this manuscript. The COPAS Biosort instrument was purchased from funds supported by National Institutes of Health Grant DK074797 (to I. H.).
PY - 2018/7/13
Y1 - 2018/7/13
N2 - Copper plays key roles in catalytic and regulatory biochemical reactions essential for normal growth, development, and health. Dietary copper deficiencies or mutations in copper homeostasis genes can lead to abnormal musculoskeletal development, cognitive disorders, and poor growth. In yeast and mammals, copper is acquired through the activities of the CTR1 family of high-affinity copper transporters. However, the mechanisms of systemic responses to dietary or tissue-specific copper deficiency remain unclear. Here, taking advantage of the animal model Caenorhabditis elegans for studying whole-body copper homeostasis, we investigated the role of a C. elegans CTR1 homolog, CHCA-1, in copper acquisition and in worm growth, development, and behavior. Using sequence homology searches, we identified 10 potential orthologs to mammalian CTR1. Among these genes, we found that chca-1, which is transcriptionally up-regulated in the intestine and hypodermis of C. elegans during copper deficiency, is required for normal growth, reproduction, and maintenance of systemic copper balance under copper deprivation. The intestinal copper transporter CUA-1 normally traffics to endosomes to sequester excess copper, and we found here that loss of chca-1 caused CUA-1 to mislocalize to the basolateral membrane under copper overload conditions. Moreover, animals lacking chca-1 exhibited significantly reduced copper avoidance behavior in response to toxic copper conditions compared with WT worms. These results establish that CHCA-1–mediated copper acquisition in C. elegans is crucial for normal growth, development, and copper-sensing behavior.
AB - Copper plays key roles in catalytic and regulatory biochemical reactions essential for normal growth, development, and health. Dietary copper deficiencies or mutations in copper homeostasis genes can lead to abnormal musculoskeletal development, cognitive disorders, and poor growth. In yeast and mammals, copper is acquired through the activities of the CTR1 family of high-affinity copper transporters. However, the mechanisms of systemic responses to dietary or tissue-specific copper deficiency remain unclear. Here, taking advantage of the animal model Caenorhabditis elegans for studying whole-body copper homeostasis, we investigated the role of a C. elegans CTR1 homolog, CHCA-1, in copper acquisition and in worm growth, development, and behavior. Using sequence homology searches, we identified 10 potential orthologs to mammalian CTR1. Among these genes, we found that chca-1, which is transcriptionally up-regulated in the intestine and hypodermis of C. elegans during copper deficiency, is required for normal growth, reproduction, and maintenance of systemic copper balance under copper deprivation. The intestinal copper transporter CUA-1 normally traffics to endosomes to sequester excess copper, and we found here that loss of chca-1 caused CUA-1 to mislocalize to the basolateral membrane under copper overload conditions. Moreover, animals lacking chca-1 exhibited significantly reduced copper avoidance behavior in response to toxic copper conditions compared with WT worms. These results establish that CHCA-1–mediated copper acquisition in C. elegans is crucial for normal growth, development, and copper-sensing behavior.
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U2 - 10.1074/jbc.RA118.003503
DO - 10.1074/jbc.RA118.003503
M3 - Article
C2 - 29784876
AN - SCOPUS:85051054375
VL - 293
SP - 10911
EP - 10925
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 28
ER -