Structure, expression and chromosomal localization of human p80-coilin gene.
Coiled bodies (CBs) are non-capsular nuclear bodies with a diameter of 0.3-1 micron and appear to be composed of coiled fibrils. Human autoantibodies to CBs recognize an 80-kD nuclear protein highly enriched in CBs, and this protein has been named p80-coilin. CBs are known to assemble and disassemble during the cell cycle, with the highest number of CBs occurring at mid to late G1 where p80-coilin is assembled into several small nuclear body-like structures. In S and G2 phases, CBs become larger and their number decreases and often they are undetectable during mitosis. Using a human autoantibody as a probe for expression cloning, we initially isolated a partial cDNA encoding p80-coilin. In this report, the 5' end of the complete cDNA for p80-coilin was obtained using the 5'-RACE (rapid amplification of cDNA ends) methodology. The size of the reconstructed full-length cDNA corresponds to the 2.7-kb mRNA detected in Northern blot analysis. The complete p80-coilin protein consists of 576 amino acids with a predicted molecular mass of 62,608. A putative p80-coilin pseudogene was also detected during the rescreening of p80-coilin cDNA. To confirm the validity of the cDNA sequence, three overlapping genomic DNA clones representing the human p80-coilin gene were selected for further analysis. The complete gene for p80-coilin contains 7 exons spanning approximately 25kb. Sequence analysis of exons 1 and 2 in genomic DNA clones confirmed the accuracy of the 5' cDNA sequence derived from the 5'-RACE procedure. Furthermore, the human p80-coilin gene was localized to chromosome 17q22-23 by fluorescence in situ hybridization.
W.M. Keck Autoimmune Disease Center, La Jolla, CA 92037.
E K
EK
Chan
S
S
Takano
L E
LE
Andrade
J C
JC
Hamel
A G
AG
Matera
D000595:Amino Acid Sequence; D001323:Autoantibodies; D001483:Base Sequence; D015152:Blotting, Northern; D002874:Chromosome Mapping; D002886:Chromosomes, Human, Pair 17; D018076:DNA, Complementary / Q000737:chemistry; D006801:Humans; D007163:Immunosorbent Techniques; D017404:In Situ Hybridization, Fluorescence; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics*; D012333:RNA, Messenger / Q000737:chemistry; D011994:Recombinant Proteins; D017421:Sequence Analysis
D001323:Autoantibodies; D018076:DNA, Complementary; D009687:Nuclear Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; C068921:p80-coilin