Molecular cloning of a novel 130-kDa cytoplasmic protein, Ankhzn, containing Ankyrin repeats hooked to a zinc finger motif.
A novel gene was trapped in mouse embryonic stem cells with a promoterless gene trap vector. Fused transcripts were isolated from the embryos by rapid amplification of cDNA ends, which were used for full-length cDNA cloning. The protein predicted from the cDNA consisting of 7143 nucleotides comprises 1184 amino acids, which was confirmed by in vitro transcription/translation assaying. An antibody against the synthesized peptide reacted with an approximate 130-kDa protein on SDS-PAGE. A search of available databases revealed that this protein is a novel protein composed of 17 ankyrin repeats hooked to a zinc finger motif, which we named Ankhzn. Ankhzn was observed on the endosomal membrane on immunoelectron microscopic analysis. Ankhzn belongs to a new subgroup of double zinc finger proteins which may be involved in vesicle or protein transport. Ankhzn mRNA and its protein were expressed ubiquitously from embryonic day 10.5 to adulthood.
Ito
K
K
Research Laboratory for Molecular Genetics, Niigata University, Niigata, 951-8510, Japan.
Ishii
N
N
Miyashita
A
A
Tominaga
K
K
Kuriyama
H
H
Maruyama
H
H
Shirai
M
M
Naito
M
M
Arakawa
M
M
Kuwano
R
R
D000595:Amino Acid Sequence; D000818:Animals; D017089:Ankyrin Repeat / Q000235:genetics*; D001483:Base Sequence; D001921:Brain / Q000196:embryology / Q000254:growth & development / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D003001:Cloning, Molecular; D003593:Cytoplasm / Q000378:metabolism* / Q000648:ultrastructure; D018076:DNA, Complementary / Q000235:genetics; D011992:Endosomes / Q000378:metabolism / Q000648:ultrastructure; D018507:Gene Expression Regulation, Developmental; D007668:Kidney / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames / Q000235:genetics; D028044:Phosphate-Binding Proteins; D012333:RNA, Messenger / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013234:Stem Cells / Q000378:metabolism; D016335:Zinc Fingers / Q000235:genetics*
C498453:Ankfy1 protein, mouse; D002352:Carrier Proteins; D018076:DNA, Complementary; D028044:Phosphate-Binding Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins