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PMID:15019990

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Citation

Mackenzie, NC, Brito, M, Reyes, AE and Allende, ML (2004) Cloning, expression pattern and essentiality of the high-affinity copper transporter 1 (ctr1) gene in zebrafish. Gene 328:113-20

Abstract

The high-affinity copper transporter 1 (Ctr1) is a highly conserved transmembrane protein that mediates the internalization of copper ions from the extracellular medium. In this study, we have isolated the zebrafish ctr1 gene. The zebrafish ctr1 cDNA encodes a protein with 69% identity to the human orthologue and shows conservation of specific amino acid residues involved in copper transport. We find only a single ctr1 gene in the zebrafish genome which maps to linkage group 5. The genomic structure of the zebrafish gene shows that it consists of five exons and that exon-intron boundaries are absolutely conserved with the mammalian ctr1 genes. Expression in embryos was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and by in situ hybridization. Zebrafish ctr1 is maternally loaded, and transcripts can be detected throughout development and in adult fish. Distribution of ctr1 message appears ubiquitous during early stages becoming restricted to the brain and ventral tissues by 24 h post fertilization (hpf). Beginning at 3 days post fertilization (dpf), expression is found mainly in the developing intestine. Specific knockdown of ctr1 by antisense morpholino oligonucleotides (MOs) causes early larval lethality. Defects include cell death in tissues where ctr1 is most heavily expressed, a finding similar to that described for a mouse knockout of mCtr1. Despite the existence of at least one other copper transport mechanism in the fish, our studies show that zebrafish ctr1 is an essential gene for development.

Links

PubMed Online version:10.1016/j.gene.2003.11.019

Keywords

Amino Acid Sequence; Animals; Base Sequence; Binding Sites/genetics; Cation Transport Proteins/genetics; Cation Transport Proteins/metabolism; Cloning, Molecular; DNA, Complementary/chemistry; DNA, Complementary/genetics; Embryo, Nonmammalian/drug effects; Embryo, Nonmammalian/metabolism; Embryonic Development; Exons; Female; Gene Expression Profiling; Gene Expression Regulation, Developmental/drug effects; Genes/genetics; Genes, Essential/genetics; In Situ Hybridization; Introns; Male; Membrane Transport Proteins/genetics; Microinjections; Molecular Sequence Data; Oligonucleotides, Antisense/pharmacology; RNA, Messenger/genetics; RNA, Messenger/metabolism; Reverse Transcriptase Polymerase Chain Reaction; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Zebrafish/embryology; Zebrafish/genetics; Zebrafish Proteins/genetics

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

DANRE:A0A2R8QTW6

enables

GO:0005375: copper ion transmembrane transporter activity

ECO:0000250: sequence similarity evidence used in manual assertion

UniProtKB:O15431

F

Seeded From UniProt

complete

DANRE:A0A2R8QTW6

involved_in

GO:0006825: copper ion transport

ECO:0000250: sequence similarity evidence used in manual assertion

UniProtKB:O15431

P

Seeded From UniProt

complete

DANRE:A2CES8

enables

GO:0005375: copper ion transmembrane transporter activity

ECO:0000250: sequence similarity evidence used in manual assertion

UniProtKB:O15431

F

Seeded From UniProt

complete

DANRE:A2CES8

involved_in

GO:0006825: copper ion transport

ECO:0000250: sequence similarity evidence used in manual assertion

UniProtKB:O15431

P

Seeded From UniProt

complete

DANRE:A7MCI3

enables

GO:0005375: copper ion transmembrane transporter activity

ECO:0000250: sequence similarity evidence used in manual assertion

UniProtKB:O15431

F

Seeded From UniProt

complete

DANRE:A7MCI3

involved_in

GO:0006825: copper ion transport

ECO:0000250: sequence similarity evidence used in manual assertion

UniProtKB:O15431

P

Seeded From UniProt

complete

DANRE:F1QKH1

enables

GO:0005375: copper ion transmembrane transporter activity

ECO:0000250: sequence similarity evidence used in manual assertion

UniProtKB:O15431

F

Seeded From UniProt

complete

DANRE:F1QKH1

involved_in

GO:0006825: copper ion transport

ECO:0000250: sequence similarity evidence used in manual assertion

UniProtKB:O15431

P

Seeded From UniProt

complete

DANRE:Q8QH07

enables

GO:0005375: copper ion transmembrane transporter activity

ECO:0000250: sequence similarity evidence used in manual assertion

UniProtKB:O15431

F

Seeded From UniProt

complete

DANRE:Q8QH07

involved_in

GO:0006825: copper ion transport

ECO:0000250: sequence similarity evidence used in manual assertion

UniProtKB:O15431

P

Seeded From UniProt

complete

See also

References

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