Dictyostelium prespore-specific gene Dp87 encodes a sorus matrix protein.
In this paper we report on the characteristics of the product of a prespore-specific gene (Dp87) of Dictyostelium discoideum. Polyclonal antibody was made against a bacterially synthesized Dp87-encoded protein fragment. Using this antibody, the product was characterized by immunochemical and immunocytological methods. It was shown that the Dp87-encoded protein is a prespore-specific protein with a molecular mass of 83 kDa, which first appears at the standing slug stage and persists in mature fruiting bodies. Western blot studies revealed the presence of an additional 81 kDa protein prior to the appearance of the 83 kDa protein from the tipped aggregate to the standing slug stage, thus indicating the former to be a precursor protein. Immunocytochemical and immunoelectron microscopic studies showed that the protein is bound to ER at the early stages of development when only the 81 kDa protein is present. At the later stages when the 83 kDa protein predominates, however, it becomes localized in prespore-specific vacuoles (PSVs) and is associated with the inner fibrous material of PSVs, but not with the peripheral membranous material. This is in contrast to spore coat proteins, which are localized in PSVs from the beginning of their appearance and associated with both structures of PSVs. In mature fruiting bodies, most Dp87 protein is localized to the interspore space (matrix) of the sori, with some left on the surface of the stalk tube. Disruptants of the Dp87 gene were also produced. Although they contained neither 81 kDa nor 83 kDa protein, they showed no phenotypic defects as compared to the parental strain.
Nakao
H
H
National Institute for Basic Biology, Okazaki, Japan.
Yamamoto
A
A
Takeuchi
I
I
Tasaka
M
M
D000818:Animals; D015153:Blotting, Western; D004023:Dictyostelium / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D005786:Gene Expression Regulation; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron; D011506:Proteins / Q000235:genetics*; D013170:Spores
D011506:Proteins