Topogenesis and sorting of synaptophysin: synthesis of a synaptic vesicle protein from a gene transfected into nonneuroendocrine cells.
Diverse nonneuroendocrine (non-NE) cells were forced to express synaptophysin (SY), the major and typical transmembrane glycoprotein of small (30-80 nm) neurotransmitter vesicles of NE cells, using microinjection of RNA synthesized in vitro from cDNA or transient and stable transfections with cDNA brought under SV40 promoter control. The glycoprotein synthesized in non-NE cells is indistinguishable from SY of NE cells and is integrated with similar, if not identical, orientation in the membranes of a specific, novel type of small cytoplasmic vesicle that structurally resembles synaptic vesicles and in which SY is the only major protein detected. A non-N-glycosylated form of SY generated by site-directed mutagenesis showed the same behavior and specific distribution in small vesicles. The results show that the information contained in this protein alone is sufficient to secure its sorting into a special type of vesicle in a heterotypic context, i.e., in the absence of other NE-specific components.
Leube
R E
RE
Institute of Cell and Tumor Biology, German Cancer Research Center, Heidelberg, Federal Republic of Germany.
Wiedenmann
B
B
Franke
W W
WW
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D005455:Fluorescent Antibody Technique; D005796:Genes; D006031:Glycosylation; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D009154:Mutation; D009368:Neoplasm Transplantation; D009419:Nerve Tissue Proteins / Q000235:genetics*; D015345:Oligonucleotide Probes; D015388:Organelles / Q000378:metabolism / Q000648:ultrastructure; D010957:Plasmids; D012313:RNA / Q000235:genetics; D051381:Rats; D015183:Restriction Mapping; D013572:Synaptic Vesicles / Q000378:metabolism*; D016708:Synaptophysin; D014158:Transcription, Genetic; D014162:Transfection
D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D015345:Oligonucleotide Probes; D016708:Synaptophysin; D012313:RNA; D004247:DNA