Molecular characterization of artemin and ferritin from Artemia franciscana.
Embryos of the brine shrimp, Artemia franciscana, exhibit remarkable resistance to physiological stress, which is temporally correlated with the presence of two proteins, one a small heat shock/alpha-crystallin protein termed p26 and the other called artemin, of unknown function. Artemin was sequenced previously by Edman degradation, and its relationship to ferritin, an iron storage protein, established. The isolation from an Artemia expressed sequence tag library of artemin and ferritin cDNAs extends this work. Artemin cDNA was found to contain an ORF of 693 nucleotides, and its deduced amino-acid sequence, except for the initiator methionine, was identical with that determined previously. Ferritin cDNA is 725 bp in length with an ORF of 516 nucleotides. Artemin amino-acid residues 32-185 are most similar to ferritin, but artemin is enriched in cysteines. The abundance of cysteines and their intramolecular spatial distribution suggest that artemin protects embryos against oxidative damage and/or that its function is redox regulated. The conserved regions in artemin and ferritin monomers are structurally similar to one another and both proteins assemble into oligomers. However, modeling of the quaternary structure indicated that artemin multimers lack the central space used for metal storage that characterizes ferritin oligomers, implying different roles for this protein. Probing of Northern blots revealed two artemin transcripts, one of 3.5 kb and another of 2.2 kb. These transcripts decreased in parallel and had almost disappeared by 16 h of development. The ferritin transcript of 0.8 kb increased slightly during reinitiation of development, then declined, and was almost completely gone by 16 h. Clearly, the loss of artemin and ferritin during embryo development is due to transcriptional regulation and proteolytic degradation of the proteins.
Chen
Tao
T
Department of Biology, Dalhousie University, Halifax, Nova Scotia, Canada.
Amons
Reinout
R
Clegg
James S
JS
Warner
Alden H
AH
MacRae
Thomas H
TH
D000595:Amino Acid Sequence; D000818:Animals; D001156:Artemia / Q000235:genetics* / Q000254:growth & development; D060829:Arthropod Proteins; D001483:Base Sequence; D002352:Carrier Proteins; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005293:Ferritins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D033862:Iron-Binding Proteins; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D011487:Protein Conformation; D016601:RNA-Binding Proteins; D040681:Structural Homology, Protein
D060829:Arthropod Proteins; D002352:Carrier Proteins; D018076:DNA, Complementary; D033862:Iron-Binding Proteins; D009419:Nerve Tissue Proteins; D016601:RNA-Binding Proteins; C490888:artemin protein, brine shrimp; D005293:Ferritins