Molecular analysis of the monomeric GTP-binding proteins of oligodendrocytes.
Vesicle transport plays an important role in the formation of myelin. Transport of proteins, including proteolipid protein and myelin associated glycoprotein, from their site of synthesis in the endoplasmic reticulum in the perikaryon of the oligodendrocytes, to myelin, takes place via carrier vesicles. The mechanisms that regulate vesicle transport in oligodendrocytes are largely unknown. The presence of monomeric GTP-binding proteins in myelin and oligodendrocytes suggested the hypothesis that these proteins participate in the regulation of vesicle transport. In an attempt to identify the Rab and Rho GTP-binding proteins present in oligodendrocytes, a cDNA library specific for these proteins was generated using a reverse transcriptase-polymerase chain reaction (RT-PCR) approach. Twelve different clones containing sequences that coded for members of the Rab and Rho families of GTP-binding proteins were isolated. This group includes Rab1, -1b, -2, -5b, -5c, -7, -8, -12, -14, -23 and Rho A. One additional clone revealed a novel cDNA sequence. Analysis of the effector loop motif indicated that this sequence encodes for a member of the Rab family. We refer to this new sequence as Rab0. Comparison of Rab0 with the most similar rat Rab sequences, Rab 14 and Rab 22, and with a recently cloned human Rab22b, showed a 71%, 72% and 94% identity, respectively. By RT-PCR analysis the Rab0 mRNA was found to be mainly expressed in oligodendrocytes and to a lesser extent in oligodendrocyte precursors, astrocytes and microglia. Moreover, the highest levels of Rab0 mRNA were observed in areas of the brain that are heavily myelinated. Rab0 mRNA was also detected in other tissues such as kidney, liver, skeletal muscle. These data provide initial evidence regarding signal transduction pathways that regulate intracellular transport in oligodendrocytes.
Burcelin
R
R
Department of Neurology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Rodriguez-Gabin
A G
AG
Charron
M J
MJ
Almazan
G
G
Larocca
J N
JN
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D019204:GTP-Binding Proteins / Q000032:analysis*; D005815:Genetic Code; D008969:Molecular Sequence Data; D009836:Oligodendroglia / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic
D012333:RNA, Messenger; D019204:GTP-Binding Proteins