Structural characterization of Dictyostelium discoideum prespore-specific gene D19 and of its product, cell surface glycoprotein PsA.
The Dictyostelium discoideum cell surface antigen PsA is a glycoprotein which first appears in the multicellular stage soon after tip formation and is selectively expressed on prespore cells. The D19 gene encodes an mRNA sequence which is highly enriched in prespore over prestalk cells in the slug stage. We have determined 81 amino acid residues of N-terminal sequence from immunoaffinity-purified PsA protein and shown this sequence to be identical to the predicted sequence of the D19 gene. There are several short repeat elements close to the C terminus, and unequal crossing-over within these is proposed to account for the size polymorphism observed in PsA protein isolated from different D. discoideum strains. The repeats are proline rich and show similarity to the C-terminal region of the D. discoideum cell adhesion molecule, contact sites A. The extreme C terminus, which is also homologous to contact sites A, is characteristic of proteins attached to the plasma membrane via a glycosyl-phosphatidylinositol link. We have marked the PsA gene by insertion of an oligonucleotide encoding an epitope of the human c-myc protein. A construct containing this gene and 990 base pairs of 5'-flanking region directed correct temporal and spatial mRNA accumulation. We found the marked PsA protein, detected with the human c-myc antibody, to be correctly localized on the surface of cells.
Early
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AE
Imperial Cancer Research Fund, Clare Hall Laboratories, Potters Bar, Hertfordshire, United Kingdom.
Williams
J G
JG
Meyer
H E
HE
Por
S B
SB
Smith
E
E
Williams
K L
KL
Gooley
A A
AA
D000595:Amino Acid Sequence; D000954:Antigens, Surface / Q000235:genetics; D001483:Base Sequence; D015815:Cell Adhesion Molecules; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004023:Dictyostelium / Q000235:genetics* / Q000502:physiology; D005656:Fungal Proteins / Q000235:genetics*; D005796:Genes; D005800:Genes, Fungal; D008562:Membrane Glycoproteins / Q000235:genetics*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D013172:Spores, Fungal / Q000502:physiology; D014158:Transcription, Genetic
D000954:Antigens, Surface; D015815:Cell Adhesion Molecules; D005656:Fungal Proteins; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D004262:DNA Restriction Enzymes