Differential expression and phosphorylation of the 74-kDa intermediate chains of cytoplasmic dynein in cultured neurons and glia.
The 74-kDa intermediate chains (IC74) of the cytoplasmic dynein complex are believed to be involved in the association of dynein with membranous organelles. While each dynein molecule is thought to have two or three IC74 subunits, at least six different IC74 protein isoforms were found in dynein from brain. Therefore we investigated the relationships of the brain cytoplasmic dynein IC74 isoforms and their association in the dynein complex at the cellular level. We found that cultured cortical neurons and glia express distinct IC74 isoforms. The IC74 isoform pattern observed in dynein from cortical neurons was generally similar to that found in dynein from adult brain, indicating that there are different populations of cytoplasmic dynein in neurons. Two IC74 isoforms were observed on two-dimensional gels of dynein from glia, while a single glial IC74 mRNA was detected. Metabolic labeling of glial dynein with 32P followed by treatment of the isolated dynein with phosphatase in vitro demonstrated that one of the glial IC74 isoforms is the product of the single glial IC74 mRNA and that the other is its phosphoisoform. A single mRNA product and its phosphoisoform are therefore sufficient for constitutive dynein function and regulation in glial cells.
Pfister
K K
KK
Cell Biology, Department School of Medicine, University of Virginia, Charlottesville 22908, USA.
Salata
M W
MW
Dillman
J F
JF
Vaughan
K T
KT
Vallee
R B
RB
Torre
E
E
Lye
R J
RJ
D017398:Alternative Splicing; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000201:enzymology*; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000201:enzymology; D003593:Cytoplasm / Q000378:metabolism; D017931:DNA Primers; D004398:Dyneins / Q000096:biosynthesis / Q000378:metabolism*; D005333:Fetus; D015870:Gene Expression; D005865:Gestational Age; D007527:Isoenzymes / Q000096:biosynthesis / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009457:Neuroglia / Q000201:enzymology*; D009474:Neurons / Q000201:enzymology*; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D051381:Rats
D017931:DNA Primers; D007527:Isoenzymes; D012333:RNA, Messenger; D004398:Dyneins