The 14-3-3 protein PAR-5 regulates the asymmetric localization of the LET-99 spindle positioning protein

Jui Ching Wu, Eugenel B. Espiritu, Lesilee S. Rose

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

PAR proteins play important roles in establishing cytoplasmic polarity as well as regulating spindle positioning during asymmetric division. However, the molecular mechanisms by which the PAR proteins generate asymmetry in different cell types are still being elucidated. Previous studies in Caenorhabditis elegans revealed that PAR-3 and PAR-1 regulate the asymmetric localization of LET-99, which in turn controls spindle positioning by affecting the distribution of the conserved force generating complex. In wild-type embryos, LET-99 is localized in a lateral cortical band pattern, via inhibition at the anterior by PAR-3 and at the posterior by PAR-1. In this report, we show that the 14-3-3 protein PAR-5 is also required for cortical LET-99 asymmetry. PAR-5 associated with LET-99 in pull-down assays, and two PAR-5 binding sites were identified in LET-99 using the yeast two-hybrid assay. Mutation of these sites abolished binding in yeast and altered LET-99 localization in vivo: LET-99 was present at the highest levels at the posterior pole of the embryo instead of a band in par-5 embryos. Together the results indicate that PAR-5 acts in a mechanism with PAR-1 to regulate LET-99 cortical localization.

Original languageEnglish (US)
Pages (from-to)288-297
Number of pages10
JournalDevelopmental Biology
Volume412
Issue number2
DOIs
StatePublished - Apr 15 2016

Fingerprint

14-3-3 Proteins
Linear Energy Transfer
Proteins
Embryonic Structures
Binding Sites
Two-Hybrid System Techniques
Caenorhabditis elegans
Yeasts
Mutation

Keywords

  • Asymmetric division
  • C. elegans
  • Embryo
  • LET-99
  • PAR-5

ASJC Scopus subject areas

  • Medicine(all)
  • Molecular Biology
  • Developmental Biology
  • Cell Biology

Cite this

The 14-3-3 protein PAR-5 regulates the asymmetric localization of the LET-99 spindle positioning protein. / Wu, Jui Ching; Espiritu, Eugenel B.; Rose, Lesilee S.

In: Developmental Biology, Vol. 412, No. 2, 15.04.2016, p. 288-297.

Research output: Contribution to journalArticle

Wu, Jui Ching ; Espiritu, Eugenel B. ; Rose, Lesilee S. / The 14-3-3 protein PAR-5 regulates the asymmetric localization of the LET-99 spindle positioning protein. In: Developmental Biology. 2016 ; Vol. 412, No. 2. pp. 288-297.
@article{6dbf15661add4cf0b0740ad1e516d53b,
title = "The 14-3-3 protein PAR-5 regulates the asymmetric localization of the LET-99 spindle positioning protein",
abstract = "PAR proteins play important roles in establishing cytoplasmic polarity as well as regulating spindle positioning during asymmetric division. However, the molecular mechanisms by which the PAR proteins generate asymmetry in different cell types are still being elucidated. Previous studies in Caenorhabditis elegans revealed that PAR-3 and PAR-1 regulate the asymmetric localization of LET-99, which in turn controls spindle positioning by affecting the distribution of the conserved force generating complex. In wild-type embryos, LET-99 is localized in a lateral cortical band pattern, via inhibition at the anterior by PAR-3 and at the posterior by PAR-1. In this report, we show that the 14-3-3 protein PAR-5 is also required for cortical LET-99 asymmetry. PAR-5 associated with LET-99 in pull-down assays, and two PAR-5 binding sites were identified in LET-99 using the yeast two-hybrid assay. Mutation of these sites abolished binding in yeast and altered LET-99 localization in vivo: LET-99 was present at the highest levels at the posterior pole of the embryo instead of a band in par-5 embryos. Together the results indicate that PAR-5 acts in a mechanism with PAR-1 to regulate LET-99 cortical localization.",
keywords = "Asymmetric division, C. elegans, Embryo, LET-99, PAR-5",
author = "Wu, {Jui Ching} and Espiritu, {Eugenel B.} and Rose, {Lesilee S.}",
year = "2016",
month = "4",
day = "15",
doi = "10.1016/j.ydbio.2016.02.020",
language = "English (US)",
volume = "412",
pages = "288--297",
journal = "Developmental Biology",
issn = "0012-1606",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - The 14-3-3 protein PAR-5 regulates the asymmetric localization of the LET-99 spindle positioning protein

AU - Wu, Jui Ching

AU - Espiritu, Eugenel B.

AU - Rose, Lesilee S.

PY - 2016/4/15

Y1 - 2016/4/15

N2 - PAR proteins play important roles in establishing cytoplasmic polarity as well as regulating spindle positioning during asymmetric division. However, the molecular mechanisms by which the PAR proteins generate asymmetry in different cell types are still being elucidated. Previous studies in Caenorhabditis elegans revealed that PAR-3 and PAR-1 regulate the asymmetric localization of LET-99, which in turn controls spindle positioning by affecting the distribution of the conserved force generating complex. In wild-type embryos, LET-99 is localized in a lateral cortical band pattern, via inhibition at the anterior by PAR-3 and at the posterior by PAR-1. In this report, we show that the 14-3-3 protein PAR-5 is also required for cortical LET-99 asymmetry. PAR-5 associated with LET-99 in pull-down assays, and two PAR-5 binding sites were identified in LET-99 using the yeast two-hybrid assay. Mutation of these sites abolished binding in yeast and altered LET-99 localization in vivo: LET-99 was present at the highest levels at the posterior pole of the embryo instead of a band in par-5 embryos. Together the results indicate that PAR-5 acts in a mechanism with PAR-1 to regulate LET-99 cortical localization.

AB - PAR proteins play important roles in establishing cytoplasmic polarity as well as regulating spindle positioning during asymmetric division. However, the molecular mechanisms by which the PAR proteins generate asymmetry in different cell types are still being elucidated. Previous studies in Caenorhabditis elegans revealed that PAR-3 and PAR-1 regulate the asymmetric localization of LET-99, which in turn controls spindle positioning by affecting the distribution of the conserved force generating complex. In wild-type embryos, LET-99 is localized in a lateral cortical band pattern, via inhibition at the anterior by PAR-3 and at the posterior by PAR-1. In this report, we show that the 14-3-3 protein PAR-5 is also required for cortical LET-99 asymmetry. PAR-5 associated with LET-99 in pull-down assays, and two PAR-5 binding sites were identified in LET-99 using the yeast two-hybrid assay. Mutation of these sites abolished binding in yeast and altered LET-99 localization in vivo: LET-99 was present at the highest levels at the posterior pole of the embryo instead of a band in par-5 embryos. Together the results indicate that PAR-5 acts in a mechanism with PAR-1 to regulate LET-99 cortical localization.

KW - Asymmetric division

KW - C. elegans

KW - Embryo

KW - LET-99

KW - PAR-5

UR - http://www.scopus.com/inward/record.url?scp=84959869578&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84959869578&partnerID=8YFLogxK

U2 - 10.1016/j.ydbio.2016.02.020

DO - 10.1016/j.ydbio.2016.02.020

M3 - Article

VL - 412

SP - 288

EP - 297

JO - Developmental Biology

JF - Developmental Biology

SN - 0012-1606

IS - 2

ER -