DOI: 10.1083/jcb.142.5.1289. The Yeast V159N Actin Mutant Reveals Roles for Actin Dynamics In Vivo

The Yeast V159N Actin Mutant Reveals Roles for Actin Dynamics In Vivo

10.1083/jcb.142.5.1289

Crossref journal-article

Rockefeller University Press

The Journal of Cell Biology (291)

Abstract

Actin with a Val 159 to Asn mutation (V159N) forms actin filaments that depolymerize slowly because of a failure to undergo a conformational change after inorganic phosphate release. Here we demonstrate that expression of this actin results in reduced actin dynamics in vivo, and we make use of this property to study the roles of rapid actin filament turnover. Yeast strains expressing the V159N mutant (act1-159) as their only source of actin have larger cortical actin patches and more actin cables than wild-type yeast. Rapid actin dynamics are not essential for cortical actin patch motility or establishment of cell polarity. However, fluid phase endocytosis is defective in act1-159 strains. act1-159 is synthetically lethal with cofilin and profilin mutants, supporting the conclusion that mutations in all of these genes impair the polymerization/ depolymerization cycle. In contrast, act1-159 partially suppresses the temperature sensitivity of a tropomyosin mutant, and the loss of cytoplasmic cables seen in fimbrin, Mdm20p, and tropomyosin null mutants, suggesting filament stabilizing functions for these actin-binding proteins. Analysis of the cables in these double-mutant cells supports a role for fimbrin in organizing cytoplasmic cables and for Mdm20p and tropomyosin in excluding cofilin from the cables.

Bibliography

Belmont, L. D., & Drubin, D. G. (1998). The Yeast V159N Actin Mutant Reveals Roles for Actin Dynamics In Vivo. The Journal of Cell Biology, 142(5), 1289â1299.

Authors 2

Lisa D. Belmont (first)
David G. Drubin (additional)

References 33 Referenced 93

10.1038/354404a0 / Nature / Requirement of yeast fimbrin for actin organization and morphogenesis in vivo by Adams (1991)
10.1083/jcb.137.2.399 / J Cell Biol / High rates of actin filament turnover in budding yeast and roles for actin in establishment and maintenance of cell polarity revealed using the actin inhibitor latrunculin-A by Ayscough (1997)
10.1002/cm.970020102 / Cell Motil / Tropomyosin binding to F-actin protects the F-actin from disassembly by brain actin-depolymerizing-factor (ADF) by Berstein (1982)
10.1016/0065-227X(90)90007-G / Adv Biophys / Actin polymerization and ATP hydrolysis by Carlier (1990)
10.1021/bi00372a001 / Biochemistry / Direct evidence for ADP-Pi-F-actin as the major intermediate in ATP-actin polymerization. Rate of dissociation of Pi from actin filaments by Carlier (1986)
10.1083/jcb.136.6.1307 / J Cell Biol / Actin depolymerizing factor (ADF/cofilin) enhances the rate of filament turnover: Implication in actin-based motility by Carlier (1997)
10.1016/0014-5793(87)81513-2 / FEBS (Fed Eur Biochem Soc) Lett / Inhibition of actin polymerization by latrunculin A by Coue (1987)
10.1073/pnas.93.9.3886 / Proc Nat Acad Sci USA / Movement of yeast cortical actin cytoskeleton visualized in vivo by Doyle (1996)
10.1083/jcb.107.6.2551 / J Cell Biol / Yeast actin-binding proteins: Evidence for a role in morphogenesis by Drubin (1988)
10.1083/jcb.113.5.1081 / J Cell Biol / Mechanism of the interaction of human platelet profilin with actin by Goldschmidt-Clermont (1981)
{'key': '2023072900032726000_B11'}
10.1083/jcb.110.1.105 / J Cell Biol / Purification of profilin from Saccharomyces cerevisiaeand analysis of profilin-deficient cells by Haarer (1990)
{'key': '2023072900032726000_B13', 'first-page': '7864', 'article-title': 'Mutational analysis of yeast profilin', 'volume': '13', 'author': 'Haarer', 'year': '1993', 'journal-title': 'Mol Cell Biol'} / Mol Cell Biol / Mutational analysis of yeast profilin by Haarer (1993)
{'key': '2023072900032726000_B14'}
10.1083/jcb.137.1.141 / J Cell Biol / The yeast gene, MDM20 is necessary for Mitochondrial Inheritance and organization of the actin cytoskeleton by Hermann (1997)
10.1038/347037a0 / Nature / Atomic structure of the actin: DNase I complex by Kabsch (1990)
10.1083/jcb.131.6.1483 / J Cell Biol / Actin filaments in yeast are unstable in the absence capping protein or fimbrin by Karpova (1995)
10.1038/40418 / Nature / Cofilin promotes rapid actin filament turnover in vivo by Lappalainen (1997)
10.1093/emboj/16.18.5520 / EMBO (Eur Mol Biol Organ) J / Essential functions and actin-binding surfaces of yeast cofilin revealed by systematic mutagenesis by Lappalainen (1997)
10.1038/314472a0 / Nature / Specific interaction between phosphatidylinositol 4,5-bisphosphate and profilactin by Lassing (1985)
10.1083/jcb.138.1.95 / J Cell Biol / In vitro reconstitution of cortical actin assembly sites in budding yeast by Lechler (1997)
10.1016/0092-8674(89)90961-6 / Cell / Disruption of the single tropomyosin gene in yeast results in the disappearance of actin cables from the cytoskeleton by Liu (1989)
10.1083/jcb.120.2.421 / J Cell Biol / Cofilin is an essential component of the yeast cortical cytoskeleton by Moon (1993)
10.1073/pnas.95.11.6181 / Proc Natl Acad Sci USA / The interaction of Arp2/3 complex with actin: Nucleation, high affinity pointed end capping, and formation of branching networks of filaments by Mullins (1998)
10.1016/0092-8674(93)90544-Z / Cell / How profilin promotes actin filament assembly in the presence of thymosin β4 by Pantaloni (1993)
10.1083/jcb.136.6.1323 / J Cell Biol / Xenopus actin depolymerizing factor/cofilin (XAC) is responsible for the turnover of actin filaments in Listeria monocytogenestails by Rosenblatt (1997)
10.1016/0955-0674(95)80051-4 / Curr Opin Cell Biol / Actin monomer binding proteins by Sun (1995)
10.1038/357257a0 / Nature / The rate of actin-based motility of intracellular Listeria monocytogenesequals the rate of actin polymerization by Theriot (1992)
10.1083/jcb.132.5.861 / J Cell Biol / Movement of cortical actin patches in yeast by Waddle (1996)
10.1016/0955-0674(94)90124-4 / Curr Opin Cell Biol / The yeast actin cytoskeleton by Welch (1994)
10.1126/science.281.5373.105 / Science / Interaction of human Arp2/3 complex and the Listeria monocytogenesActA protein in actin filament nucleation by Welch (1998)
10.1093/genetics/132.2.337 / Genetics / Systematic mutational analysis of the yeast ACT1 gene by Wertman (1992)
10.1016/S0960-9822(06)00223-5 / Curr Biol / The complex containing actin-related proteins Arp2 and Arp3 is required for the motility and integrity of yeast actin patches by Winter (1997)

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 12:47 p.m.)
Deposited	2 years, 1 month ago (July 28, 2023, 8:04 p.m.)
Indexed	9 months, 1 week ago (Nov. 19, 2024, 10:41 a.m.)
Issued	26 years, 11 months ago (Sept. 7, 1998)
Published	26 years, 11 months ago (Sept. 7, 1998)
Published Online	26 years, 11 months ago (Sept. 7, 1998)
Published Print	26 years, 11 months ago (Sept. 7, 1998)

Funders 0

None

BibTeX

@article{Belmont_1998, title={The Yeast V159N Actin Mutant Reveals Roles for Actin Dynamics In Vivo}, volume={142}, ISSN={1540-8140}, url={http://dx.doi.org/10.1083/jcb.142.5.1289}, DOI={10.1083/jcb.142.5.1289}, number={5}, journal={The Journal of Cell Biology}, publisher={Rockefeller University Press}, author={Belmont, Lisa D. and Drubin, David G.}, year={1998}, month=sep, pages={1289–1299} }

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  "URL": "http://dx.doi.org/10.1083/jcb.142.5.1289",
  "relation": {},
  "ISSN": [
    "0021-9525",
    "1540-8140"
  ],
  "subject": [],
  "published": {
    "date-parts": [
      [
        1998,
        9,
        7
      ]
    ]
  }
}