Expression study on D123 gene product: evidence for high positivity in testis.
A novel 44-kDa gene product (D123) has been proposed as necessary for S-phase entry of the cell cycle: a point mutation resulted in a temperature-sensitive arrest in G1-phase. From human fibrosarcoma cDNA library, we have isolated an identical gene and studied its sequence and mRNA and protein expression. Compared with D123, three nucleotide differences within the human coding sequence, plus others, result in a change of two amino acids. A partial sequence similarity has been found with a yeast gene of unknown function. The protein has several potential phosphorylation sites, is highly hydrophilic, and may be highly structured in alpha-helix. The mRNA is abundantly expressed by a variety of normal and transformed cells and by all tissues examined, being most highly expressed in testis. Specific antibodies, raised against a rhD123 polypeptide, recognize a major 42- to 44-kDa molecule in crude extract of various human cell lines. Immunohistochemistry reveals that D123 protein is not homogeneously expressed: it is detected, often in granular vescicles, in the cytoplasm of some epithelial, stromal, and sperm cells and in varicosities lining nervous fibers, while it appears to be absent in nuclei, endothelial, and smooth muscle cells. The precise link between cytoplasmic occurrence of D123 and cell cycle progression still remains to be clarified.
Medical School, Department of Biological Chemistry, University of Padua, viale G. Colombo 3, Padua, 35121, Italy. onisto@civ.bio.unipd.it
M
M
Onisto
P
P
Zeilante
P
P
Scannapieco
D
D
Pellati
M
M
Pozza
C
C
Caenazzo
A
A
Negro
S
S
Garbisa
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D018797:Cell Cycle Proteins; D002460:Cell Line; D002461:Cell Line, Transformed; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D015870:Gene Expression / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000032:analysis / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013223:Statistics as Topic; D013737:Testis / Q000737:chemistry; D014018:Tissue Distribution; D014407:Tumor Cells, Cultured
D000596:Amino Acids; C484374:Cdc123 protein, rat; D018797:Cell Cycle Proteins; D018076:DNA, Complementary; D011506:Proteins; D012333:RNA, Messenger; C098828:CDC123 protein, human