Abstract
A previously uncharacterized yeast gene (YER016w) that we have named BIM1 (binding to microtubules) was obtained from a two-hybrid screen of a yeast cDNA library using as bait the entire coding sequence of TUB1 (encoding alpha-tubulin). Deletion of BIM1 results in a strong bilateral karyogamy defect, hypersensitivity to benomyl, and aberrant spindle behavior, all phenotypes associated with mutations affecting microtubules in yeast, and inviability at extreme temperatures (i.e., >/=37 degrees C or </=14 degrees C). Overexpression of BIM1 in wild-type cells is lethal. A fusion of Bim1p with green fluorescent protein that complements the bim1Delta phenotypes allows visualization in vivo of both intranuclear spindles and extranuclear microtubules in otherwise wild-type cells. A bim1 deletion displays synthetic lethality with deletion alleles of bik1, num1, and bub3 as well as a limited subset of tub1 conditional-lethal alleles. A systematic study of 51 tub1 alleles suggests a correlation between specific failure to interact with Bim1p in the two-hybrid assay and synthetic lethality with the bim1Delta allele. The sequence of BIM1 shows substantial similarity to sequences from organisms across the evolutionary spectrum. One of the human homologues, EB1, has been reported previously as binding APC, itself a microtubule-binding protein and the product of a gene implicated in the etiology of human colon cancer.
MeSH Terms
Alleles
Amino Acid Sequence
Cell Cycle
Cell Cycle Proteins/chemistry,genetics,metabolism
Cell Nucleus/chemistry,metabolism
Cell Size
Cytoskeleton/chemistry,genetics,metabolism
Fungal Proteins/chemistry,genetics,metabolism
Genes, Fungal/genetics,physiology
Genes, Lethal/genetics
Humans
Microtubule Proteins/chemistry,genetics,metabolism
Microtubule-Associated Proteins/chemistry
Microtubules/chemistry,genetics,metabolism
Molecular Sequence Data
Mutation/genetics
Nuclear Proteins/chemistry,genetics,metabolism
Phenotype
Protein Binding
Recombinant Fusion Proteins/chemistry,genetics,metabolism
Saccharomyces cerevisiae/cytology,genetics
Saccharomyces cerevisiae Proteins
Spindle Apparatus/chemistry,metabolism
Tubulin/chemistry,genetics,metabolism
Two-Hybrid System Techniques
Chemicals
BIM1 protein, S cerevisiae
Cell Cycle Proteins
EB1 microtubule binding proteins
Fungal Proteins
Microtubule Proteins
Microtubule-Associated Proteins
Nuclear Proteins
Recombinant Fusion Proteins
Saccharomyces cerevisiae Proteins
Tubulin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Schwartz K
Department of Genetics, Stanford University School of Medicine, Stanford, California 94305, USA.
Richards K
Botstein D
References (37)
37 references, click to expand
-
Transformation of intact yeast cells treated with alkali cations.
J Bacteriol. 1983 Jan;153(1):163-8
PMID: 6336730
-
MHP1, an essential gene in Saccharomyces cerevisiae required for microtubule function.
J Cell Biol. 1996 Dec;135(5):1323-39
PMID: 8947554
-
Isolation of the beta-tubulin gene from yeast and demonstration of its essential function in vivo.
Cell. 1983 May;33(1):211-9
PMID: 6380751
-
A yeast mutant temperature-sensitive for mitochondrial assembly is deficient in a mitochondrial protease activity that cleaves imported precursor polypeptides.
EMBO J. 1985 Aug;4(8):2069-74
PMID: 2866092
-
Genetically essential and nonessential alpha-tubulin genes specify functionally interchangeable proteins.
Mol Cell Biol. 1986 Nov;6(11):3722-33
PMID: 3540600
-
KAR1, a gene required for function of both intranuclear and extranuclear microtubules in yeast.
Cell. 1987 Mar 27;48(6):1047-60
PMID: 3030557
-
Two genes required for cell fusion during yeast conjugation: evidence for a pheromone-induced surface protein.
Mol Cell Biol. 1987 Jul;7(7):2316-28
PMID: 3302672
-
Improved tools for biological sequence comparison.
Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444-8
PMID: 3162770
-
Diverse effects of beta-tubulin mutations on microtubule formation and function.
J Cell Biol. 1988 Jun;106(6):1997-2010
PMID: 3290223
-
Isolation and characterization of conditional-lethal mutations in the TUB1 alpha-tubulin gene of the yeast Saccharomyces cerevisiae.
Genetics. 1988 Nov;120(3):681-95
PMID: 3066684
-
KAR3, a kinesin-related gene required for yeast nuclear fusion.
Cell. 1990 Mar 23;60(6):1029-41
PMID: 2138512
-
Yeast Saccharomyces cerevisiae selectable markers in pUC18 polylinkers.
Yeast. 1990 Sep-Oct;6(5):363-6
PMID: 2220072
-
The ADE2 gene from Saccharomyces cerevisiae: sequence and new vectors.
Gene. 1990 Oct 30;95(1):91-8
PMID: 2253890
-
BIK1, a protein required for microtubule function during mating and mitosis in Saccharomyces cerevisiae, colocalizes with tubulin.
J Cell Biol. 1990 Dec;111(6 Pt 1):2573-86
PMID: 2277073
-
S. cerevisiae genes required for cell cycle arrest in response to loss of microtubule function.
Cell. 1991 Aug 9;66(3):507-17
PMID: 1651171
-
Identification and characterization of the familial adenomatous polyposis coli gene.
Cell. 1991 Aug 9;66(3):589-600
PMID: 1651174
-
Nuclear migration in Saccharomyces cerevisiae is controlled by the highly repetitive 313 kDa NUM1 protein.
Mol Gen Genet. 1991 Nov;230(1-2):277-87
PMID: 1745235
-
Yeast proteins associated with microtubules in vitro and in vivo.
Mol Biol Cell. 1992 Jan;3(1):29-47
PMID: 1348005
-
Two Saccharomyces cerevisiae kinesin-related gene products required for mitotic spindle assembly.
J Cell Biol. 1992 Jul;118(1):109-20
PMID: 1618897
-
Kinesin-related proteins required for assembly of the mitotic spindle.
J Cell Biol. 1992 Jul;118(1):95-108
PMID: 1618910
-
Wild-type but not mutant APC associates with the microtubule cytoskeleton.
Cancer Res. 1994 Jul 15;54(14):3672-5
PMID: 8033082
-
The APC gene product associates with microtubules in vivo and promotes their assembly in vitro.
Cancer Res. 1994 Jul 15;54(14):3676-81
PMID: 8033083
-
Mapping actin surfaces required for functional interactions in vivo.
J Cell Biol. 1994 Jul;126(2):423-32
PMID: 8034743
-
The two-hybrid system: an assay for protein-protein interactions.
Trends Genet. 1994 Aug;10(8):286-92
PMID: 7940758
-
STU1, a suppressor of a beta-tubulin mutation, encodes a novel and essential component of the yeast mitotic spindle.
J Cell Biol. 1994 Dec;127(6 Pt 2):1973-84
PMID: 7806575
-
Improved green fluorescence.
Nature. 1995 Feb 23;373(6516):663-4
PMID: 7854443
-
Defining protein interactions with yeast actin in vivo.
Nat Struct Biol. 1995 Jan;2(1):28-35
PMID: 7719850
-
A role of Sep1 (= Kem1, Xrn1) as a microtubule-associated protein in Saccharomyces cerevisiae.
EMBO J. 1995 Mar 15;14(6):1057-66
PMID: 7720696
-
Green fluorescent protein. The green revolution.
Curr Biol. 1995 Mar 1;5(3):262-4
PMID: 7780736
-
APC binds to the novel protein EB1.
Cancer Res. 1995 Jul 15;55(14):2972-7
PMID: 7606712
-
Two microtubule-associated proteins required for anaphase spindle movement in Saccharomyces cerevisiae.
J Cell Biol. 1995 Sep;130(6):1373-85
PMID: 7559759
-
Yeast Num1p associates with the mother cell cortex during S/G2 phase and affects microtubular functions.
J Cell Biol. 1995 Nov;131(4):1003-14
PMID: 7490278
-
Precise gene disruption in Saccharomyces cerevisiae by double fusion polymerase chain reaction.
Yeast. 1995 Oct;11(13):1275-80
PMID: 8553698
-
Movement of yeast cortical actin cytoskeleton visualized in vivo.
Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):3886-91
PMID: 8632984
-
The adenomatous polyposis coli tumor suppressor protein localizes to plasma membrane sites involved in active cell migration.
J Cell Biol. 1996 Jul;134(1):165-79
PMID: 8698812
-
Gene identification using the yeast two-hybrid system.
Methods Enzymol. 1996;273:331-47
PMID: 8791622
-
Structural rearrangements of tubulin and actin during the cell cycle of the yeast Saccharomyces.
J Cell Biol. 1984 Mar;98(3):922-33
PMID: 6365930