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PMID: 8744947 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

p150Glued, the largest subunit of the dynactin complex, is nonessential in Neurospora but required for nuclear distribution.

Molecular biology of the cell ·Vol. 7 ·No. 5 ·1996-05-00 ·Pages 731-42

Tinsley JH, Minke PF, Bruno KS, Plamann M

Abstract

Dynactin is a multisubunit complex that is required for cytoplasmic dynein, a minus-end-directed, microtubule-associated motor, to efficiently transport vesicles along microtubules in vitro. p150Glued, the largest subunit of dynactin, has been identified in vertebrates and Drosophila and recently has been shown to interact with cytoplasmic dynein intermediate chains in vitro. The mechanism by which dynactin facilitates cytoplasmic dynein-dependent vesicle transport is unknown. We have devised a genetic screen for cytoplasmic dynein/dynactin mutants in the filamentous fungus Neurospora crassa. In this paper, we report that one of these mutants, ro-3, defines a gene encoding an apparent homologue of p150Glued, and we provide genetic evidence that cytoplasmic dynein and dynactin interact in vivo. The major structural features of vertebrate and Drosophila p150Glued, a microtubule-binding site at the N-terminus and two large alpha-helical coiled-coil regions contained within the distal two-thirds of the polypeptide, are conserved in Ro3. Drosophila p150Glued is essential for viability; however, ro-3 null mutants are viable, indicating that dynactin is not an essential complex in N. crassa. We show that N. crassa cytoplasmic dynein and dynactin mutants have abnormal nuclear distribution but retain the ability to organize cytoplasmic microtubules and actin in anucleate hyphae.

MeSH Terms
Actins/physiology Amino Acid Sequence Base Sequence Cell Survival Cloning, Molecular Dynactin Complex Dyneins/metabolism Genes, Fungal Genetic Complementation Test Immunohistochemistry Microtubule Proteins/chemistry,genetics,physiology Microtubule-Associated Proteins Microtubules/physiology Molecular Sequence Data Neurospora crassa/genetics,physiology Polymorphism, Restriction Fragment Length Restriction Mapping Sequence Homology, Amino Acid
Chemicals
Actins Dynactin Complex Microtubule Proteins Microtubule-Associated Proteins Dyneins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Tinsley J H
Department of Biology, Texas A&M University, College Station 77843-3258, USA.
Minke P F
Bruno K S
Plamann M
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.7.5.731
Published
1996-05-00
Pages
731-42
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC275926
Subset
IM
Grants
NIGMS NIH HHS · GM-51217 · United States
Databases
GENBANK
L48661
External Links
PubMed source DOI: 10.1091/mbc.7.5.731 PMC Full Text View journal details
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