Tissue-specific processing and polarized compartmentalization of clone-produced cholinesterase in microinjected Xenopus oocytes.
1. To approach the involvement of tissue-specific elements in the compartmentalization of ubiquitous polymorphic proteins, immunohistochemical methods were used to analyze the localization of butyrylcholinesterase (BuChE) in Xenopus oocytes microinjected with synthetic BuChEmRNA alone and in combination with tissue-extracted mRNAs. 2. When injected alone BuChEmRNA efficiently directed the synthesis of small membrane-associated accumulations localized principally on the external surface of the oocyte's animal pole. Tunicamycin blocked the appearance of such accumulations, suggesting that glycosylation is involved in the transport of nascent BuChE molecules to the oocyte's surface. Coinjection with brain or muscle mRNA, but not liver mRNA, facilitated the formation of pronounced, tissue-characteristic BuChE aggregates. 3. These findings implicate tissue-specific mRNAs in the assembly of the clone-produced protein and in its nonuniform distribution in the oocyte membrane or extracellular material.
Dreyfus
P A
PA
Department of Biological Chemistry, Hebrew University, Jerusalem, Israel.
Seidman
S
S
Pincon-Raymond
M
M
Murawsky
M
M
Rieger
F
F
Schejter
E
E
Zakut
H
H
Soreq
H
H
D000818:Animals; D002091:Butyrylcholinesterase / Q000096:biosynthesis* / Q000235:genetics; D002462:Cell Membrane / Q000201:enzymology; D002802:Cholinesterases / Q000096:biosynthesis*; D003001:Cloning, Molecular; D005455:Fluorescent Antibody Technique; D007150:Immunohistochemistry; D008845:Microinjections; D008854:Microscopy, Electron; D009865:Oocytes / Q000201:enzymology* / Q000648:ultrastructure; D009928:Organ Specificity; D012333:RNA, Messenger / Q000235:genetics; D013347:Subcellular Fractions / Q000201:enzymology; D014158:Transcription, Genetic; D014981:Xenopus
D012333:RNA, Messenger; D002091:Butyrylcholinesterase; D002802:Cholinesterases