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Alf1p, a CLIP-170 Domain-containing Protein, Is Functionally and Physically Associated with α-Tubulin

Journal Article · · Journal of Cell Biology
 [1];  [2];  [3];  [4]
  1. Stanford Univ., CA (United States). Dept. of Biological Sciences; DOE/OSTI
  2. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Life Science Division; Univ. of California, Berkeley, CA (United States). Molecular and Cell Biology Dept.
  3. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Life Science Division
  4. Stanford Univ., CA (United States). Dept. of Biological Sciences
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Tubulin is a heterodimer of α- and β-tubulin polypeptides. Assembly of the tubulin heterodimer in vitro requires the CCT chaperonin complex, and a set of five proteins referred to as the tubulin cofactors (Tian, F., Y. Huang, H. Rommelaere, J. Vandekerckhove, C. Ampe, and N.J. Cowan. 1996. Cell. 86:287–296; Tian, G., S.A. Lewis, B. Feierbach, T. Stearns, H. Rommelaere, C. Ampe, and N.J. Cowan. 1997. J. Cell Biol. 138:821–832). We report the characterization of Alf1p, the yeast ortholog of mammalian cofactor B. Alf1p interacts with α-tubulin in both two-hybrid and immunoprecipitation assays. Alf1p and cofactor B contain a single CLIP-170 domain, which is found in several microtubule-associated proteins. Mutation of the CLIP-170 domain in Alf1p disrupts the interaction with α-tubulin. Mutations in α-tubulin that disrupt the interaction with Alf1p map to a domain on the cytoplasmic face of α-tubulin; this domain is distinct from the region of interaction between α-tubulin and β-tubulin. Alf1p-green fluorescent protein (GFP) is able to associate with microtubules in vivo, and this localization is abolished either by mutation of the CLIP-170 domain in Alf1p, or by mutation of the Alf1p-binding domain in α-tubulin. Analysis of double mutants constructed between null alleles of ALF1 and PAC2, which encodes the other yeast α-tubulin cofactor, suggests that Alf1p and Pac2p act in the same pathway leading to functional α-tubulin. The phenotype of overexpression of ALF1 suggests that Alf1p can act to sequester α-tubulin from interaction with β-tubulin, raising the possibility that it plays a regulatory role in the formation of the tubulin heterodimer.
Research Organization:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division
Grant/Contract Number:
SC0012704
OSTI ID:
1625111
Journal Information:
Journal of Cell Biology, Journal Name: Journal of Cell Biology Journal Issue: 1 Vol. 144; ISSN 0021-9525
Publisher:
Rockefeller University PressCopyright Statement
Country of Publication:
United States
Language:
English

References (54)

Function in protein folding of TRiC, a cytosolic ring complex containing TCP-1 and structurally related subunits. journal December 1992
CLIP-170 links endocytic vesicles to microtubules journal September 1992
A cytoplasmic chaperonin that catalyzes β-actin folding journal June 1992
Basic local alignment search tool journal October 1990
TCP1 complex is a molecular chaperone in tubulin biogenesis journal July 1992
Lambda YES: a multifunctional cDNA expression vector for the isolation of genes by complementation of yeast and Escherichia coli mutations. journal March 1991
The t-complex polypeptide 1 complex is a chaperonin for tubulin and actin in vivo. journal October 1993
BIK1, a protein required for microtubule function during mating and mitosis in Saccharomyces cerevisiae, colocalizes with tubulin. journal December 1990
Chaperonin-mediated folding of vertebrate actin-related protein and gamma-tubulin journal September 1993
Mapping actin surfaces required for functional interactions in vivo. journal July 1994
The Mammalian γ-Tubulin Complex Contains Homologues of the Yeast Spindle Pole Body Components Spc97p and Spc98p journal May 1998
Rat monoclonal antitubulin antibodies derived by using a new nonsecreting rat cell line journal June 1982
BIM1 Encodes a Microtubule-binding Protein in Yeast journal December 1997
Saccharomyces cerevisiae genes required in the absence of the CIN8-encoded spindle motor act in functionally diverse mitotic pathways. journal June 1997
Unlinked noncomplementation: isolation of new conditional-lethal mutations in each of the tubulin genes of Saccharomyces cerevisiae. journal June 1988
Yeast mutants sensitive to antimicrotubule drugs define three genes that affect microtubule function. journal February 1990
Saccharomyces cerevisiae PAC2 Functions With CIN1, 2 and 4 in a Pathway Leading to Normal Microtubule Stability journal July 1997
The role of molecular chaperones in protein folding. journal December 1995
The retinoblastoma protein associates with the protein phosphatase type 1 catalytic subunit. journal April 1993
Principles that Govern the Folding of Protein Chains journal July 1973
Many random sequences functionally replace the secretion signal sequence of yeast invertase journal January 1987
Behavior of spindles and spindle plaques in the cell cycle and conjugation of Saccharomyces cerevisiae. journal January 1975
Regulation of Tubulin Levels and Microtubule Assembly in Saccharomyces cerevisiae: Consequences of Altered Tubulin Gene Copy Number journal October 1990
Phenotypic consequences of tubulin overproduction in Saccharomyces cerevisiae: differences between alpha-tubulin and beta-tubulin. journal October 1990
Two Cofactors and Cytoplasmic Chaperonin Are Required for the Folding of α- and β-Tubulin journal April 1993
Two functional alpha-tubulin genes of the yeast Saccharomyces cerevisiae encode divergent proteins journal November 1986
Dominant effects of tubulin overexpression in Saccharomyces cerevisiae. journal March 1989
Molecular characterization of two functional domains of CLIP-170 in vivo journal July 1994
Review: The Cct eukaryotic chaperonin subunits of Saccharomyces cerevisiae and other yeasts journal May 1996
CLIP-170 links endocytic vesicles to microtubules journal September 1992
A cytoplasmic chaperonin that catalyzes β-actin folding journal June 1992
Rbl2p, a yeast protein that binds to β-tubulin and participates in microtubule function in vivo journal August 1995
FACS-optimized mutants of the green fluorescent protein (GFP) journal January 1996
Basic local alignment search tool journal October 1990
Chapter 19 Fluorescence Microscopy Methods for Yeast book January 1989
Pathway Leading to Correctly Folded β-Tubulin journal July 1996
Structure of the αβ tubulin dimer by electron crystallography journal January 1998
Defining protein interactions with yeast actin in vivo journal January 1995
Three-dimensional ultrastructural analysis of the Saccharomyces cerevisiae mitotic spindle journal June 1995
Analysis of Tub4p, a yeast gamma-tubulin-like protein: implications for microtubule-organizing center function. journal July 1996
Tubulin Subunits Exist in an Activated Conformational State Generated and Maintained by Protein Cofactors journal August 1997
An α-Tubulin Mutant Destabilizes the Heterodimer: Phenotypic Consequences and Interactions with Tubulin-binding Proteins journal September 1998
Essential role of tubulin-folding cofactor D in microtubule assembly and its association with microtubules in fission yeast journal February 1998
Systematic mutational analysis of the yeast ACT1 gene. journal October 1992
High-resolution epitope mapping of hGH-receptor interactions by alanine-scanning mutagenesis journal June 1989
Behavior of spindles and spindle plaques in the cell cycle and conjugation of Saccharomyces cerevisiae. journal January 1975
Chromosome instability mutants of Saccharomyces cerevisiae that are defective in microtubule-mediated processes. journal January 1990
Regulation of tubulin levels and microtubule assembly in Saccharomyces cerevisiae: consequences of altered tubulin gene copy number. journal October 1990
Phenotypic consequences of tubulin overproduction in Saccharomyces cerevisiae: differences between alpha-tubulin and beta-tubulin. journal October 1990
Two cofactors and cytoplasmic chaperonin are required for the folding of alpha- and beta-tubulin. journal April 1993
Formation and Function of the Rbl2p–β-Tubulin Complex journal March 1998
Two functional alpha-tubulin genes of the yeast Saccharomyces cerevisiae encode divergent proteins. journal November 1986
Genetically essential and nonessential alpha-tubulin genes specify functionally interchangeable proteins. journal November 1986
Dominant effects of tubulin overexpression in Saccharomyces cerevisiae. journal March 1989

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A unique cytoskeleton-associated protein in Cryptococcus neoformans journal April 2018
Adp Ribosylation Factor-like Protein 2 (Arl2) Regulates the Interaction of Tubulin-Folding Cofactor D with Native Tubulin journal May 2000
Expression of Cd34 and Myf5 Defines the Majority of Quiescent Adult Skeletal Muscle Satellite Cells journal December 2000
Myogenic cell proliferation and generation of a reversible tumorigenic phenotype are triggered by preirradiation of the recipient site journal May 2002
The dual role of fission yeast Tbc1/cofactor C orchestrates microtubule homeostasis in tubulin folding and acts as a GAP for GTPase Alp41/Arl2 journal June 2013
Revisiting the tubulin cofactors and Arl2 in the regulation of soluble αβ-tubulin pools and their effect on microtubule dynamics journal February 2017
Tubulin cofactors and Arl2 are cage-like chaperones that regulate the soluble αβ-tubulin pool for microtubule dynamics journal July 2015
Autoinhibition of TBCB regulates EB1-mediated microtubule dynamics journal September 2012
The novel microtubule-associated CAP-glycine protein Cgp1 governs growth, differentiation, and virulence of Cryptococcus neoformans journal February 2018
Drosophila dTBCE recruits tubulin around chromatin to promote mitotic spindle assembly preprint January 2020

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CLIP-170
Saccharomyces cerevisiae
cell biology
chaperonin
microtubule
tubulin