A dorso-ventral asymmetry in the embryonic retina defined by protein conformation.
In a search for determinants of retinotopic specification we previously identified an antigen in the dorsal embryonic retina as a protein called the 68-kDa laminin receptor. A dorso-ventral asymmetry in a laminin receptor seemed consistent with the known responsiveness of embryonic optic axons to laminin, but there were three peculiar points. (i) The molecular mass of this presumed laminin receptor in immunoblots is not 68 kDa but 43 kDa, and the molecular mass of the protein deduced from the mRNA is only 33 kDa. (ii) The antigen does not have the localization expected of a receptor for the extracellular matrix: the antibodies label mainly a granular cytoplasmic antigen in dorsal retina; an additional sparse cell-surface antigen present on a few cells does not show a dorso-ventral asymmetry. (iii) Despite the pronounced dorso-ventral difference seen immunohistochemically, in immunoblots the 43-kDa protein (p40) is evenly distributed throughout the retina. Here we show that (i) native p40 and in vitro-translated gene product are indistinguishable and their anomalous migration in denaturing gels probably is due to low pI; (ii) p40 is bound in a Mg2(+)-dependent manner to large cytoplasmic complexes that appear to include ribosomes; and (iii) there is a labile conformational difference in p40 between dorsal and ventral retina: dorsally it is more accessible to proteolysis, suggesting a more open conformation. In conjunction with the recent hypothesis that p40 constitutes a translation initiation factor (D. Auth and G. Brawerman, personal communication), these observations point to a dorso-ventral asymmetry in some aspect of protein translation, which in turn may set up differences in recognition factors on retinal growth cones.
McCaffery
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P
Department of Neurobiology, Harvard Medical School, Boston, MA 02115.
Neve
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RL
Dräger
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UC
D000818:Animals; D000911:Antibodies, Monoclonal; D001369:Axons / Q000502:physiology; D002460:Cell Line; D005455:Fluorescent Antibody Technique; D007797:Laminin / Q000502:physiology; D008970:Molecular Weight; D014176:Protein Biosynthesis; D011487:Protein Conformation; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011946:Receptors, Antigen / Q000032:analysis*; D011971:Receptors, Immunologic / Q000032:analysis* / Q000235:genetics / Q000302:isolation & purification; D017474:Receptors, Laminin; D012160:Retina / Q000196:embryology*
D000911:Antibodies, Monoclonal; D007797:Laminin; D012333:RNA, Messenger; D011946:Receptors, Antigen; D011971:Receptors, Immunologic; D017474:Receptors, Laminin