Critical role for the EB1 and APC interaction in the regulation of microtubule polymerization

Masafumi Nakamura, Xiao Zhen Zhou, Kun Ping Lu

Research output: Contribution to journalArticle

150 Citations (Scopus)

Abstract

Human EB1 was originally cloned as a protein that interacts with the COOH terminus of adenomatous polyposis coli (APC) [1]. Interestingly, this interaction is often disrupted in colon cancer, due to mutations in APC. EB1 also interacts with the plus-ends of microtubules and targets APC to microtubule tips [2-6]. Since APC is detected on the kinetochores of chromosomes, it has been hypothesized that the EB1-APC interaction connects microtubule spindles to the kinetochores and regulates microtubule stability [7-9]. In yeast, EB1 regulates microtubule dynamics [6, 10, 11], and its binding domain in APC may be conserved in Kar9, an EB1 binding protein involved in the microtubule-capturing mechanism [12-14]. These results suggest that the interaction of EB1 and APC is important and may be conserved. However, it is largely unknown whether the EB1-APC interaction affects microtubule dynamics. Here, we show that EB1 potently promotes microtubule polymerization in vitro and in permeabilized cells, but, surprisingly, only in the presence of the COOH-terminal EB1 binding domain of APC (C-APC). Significantly, this C-APC activity is abolished by phosphorylation, which also disrupts its ability to bind to EB1. Furthermore, yeast EB1 protein effectively substitutes for the human protein but also requires C-APC in promoting microtubule polymerization. Finally, C-APC is able to promote microtubule polymerization when stably expressed in APC mutant cells, demonstrating the ability of C-APC to promote microtubule assembly in vivo. Thus, the interaction between EB1 and APC plays an essential role in the regulation of microtubule polymerization, and a similar mechanism may be conserved in yeast.

Original languageEnglish
Pages (from-to)1062-1067
Number of pages6
JournalCurrent Biology
Volume11
Issue number13
DOIs
Publication statusPublished - Jul 10 2001
Externally publishedYes

Fingerprint

Adenomatous Polyposis Coli
Microtubules
Polymerization
polymerization
microtubules
Yeast
Phosphorylation
Chromosomes
kinetochores
Carrier Proteins
Proteins
yeasts
Kinetochores
proteins
Yeasts
colorectal neoplasms
Fungal Proteins
binding proteins
phosphorylation

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Critical role for the EB1 and APC interaction in the regulation of microtubule polymerization. / Nakamura, Masafumi; Zhou, Xiao Zhen; Lu, Kun Ping.

In: Current Biology, Vol. 11, No. 13, 10.07.2001, p. 1062-1067.

Research output: Contribution to journalArticle

@article{6814060024e44f88a0d6d2179447348b,
title = "Critical role for the EB1 and APC interaction in the regulation of microtubule polymerization",
abstract = "Human EB1 was originally cloned as a protein that interacts with the COOH terminus of adenomatous polyposis coli (APC) [1]. Interestingly, this interaction is often disrupted in colon cancer, due to mutations in APC. EB1 also interacts with the plus-ends of microtubules and targets APC to microtubule tips [2-6]. Since APC is detected on the kinetochores of chromosomes, it has been hypothesized that the EB1-APC interaction connects microtubule spindles to the kinetochores and regulates microtubule stability [7-9]. In yeast, EB1 regulates microtubule dynamics [6, 10, 11], and its binding domain in APC may be conserved in Kar9, an EB1 binding protein involved in the microtubule-capturing mechanism [12-14]. These results suggest that the interaction of EB1 and APC is important and may be conserved. However, it is largely unknown whether the EB1-APC interaction affects microtubule dynamics. Here, we show that EB1 potently promotes microtubule polymerization in vitro and in permeabilized cells, but, surprisingly, only in the presence of the COOH-terminal EB1 binding domain of APC (C-APC). Significantly, this C-APC activity is abolished by phosphorylation, which also disrupts its ability to bind to EB1. Furthermore, yeast EB1 protein effectively substitutes for the human protein but also requires C-APC in promoting microtubule polymerization. Finally, C-APC is able to promote microtubule polymerization when stably expressed in APC mutant cells, demonstrating the ability of C-APC to promote microtubule assembly in vivo. Thus, the interaction between EB1 and APC plays an essential role in the regulation of microtubule polymerization, and a similar mechanism may be conserved in yeast.",
author = "Masafumi Nakamura and Zhou, {Xiao Zhen} and Lu, {Kun Ping}",
year = "2001",
month = "7",
day = "10",
doi = "10.1016/S0960-9822(01)00297-4",
language = "English",
volume = "11",
pages = "1062--1067",
journal = "Current Biology",
issn = "0960-9822",
publisher = "Cell Press",
number = "13",

}

TY - JOUR

T1 - Critical role for the EB1 and APC interaction in the regulation of microtubule polymerization

AU - Nakamura, Masafumi

AU - Zhou, Xiao Zhen

AU - Lu, Kun Ping

PY - 2001/7/10

Y1 - 2001/7/10

N2 - Human EB1 was originally cloned as a protein that interacts with the COOH terminus of adenomatous polyposis coli (APC) [1]. Interestingly, this interaction is often disrupted in colon cancer, due to mutations in APC. EB1 also interacts with the plus-ends of microtubules and targets APC to microtubule tips [2-6]. Since APC is detected on the kinetochores of chromosomes, it has been hypothesized that the EB1-APC interaction connects microtubule spindles to the kinetochores and regulates microtubule stability [7-9]. In yeast, EB1 regulates microtubule dynamics [6, 10, 11], and its binding domain in APC may be conserved in Kar9, an EB1 binding protein involved in the microtubule-capturing mechanism [12-14]. These results suggest that the interaction of EB1 and APC is important and may be conserved. However, it is largely unknown whether the EB1-APC interaction affects microtubule dynamics. Here, we show that EB1 potently promotes microtubule polymerization in vitro and in permeabilized cells, but, surprisingly, only in the presence of the COOH-terminal EB1 binding domain of APC (C-APC). Significantly, this C-APC activity is abolished by phosphorylation, which also disrupts its ability to bind to EB1. Furthermore, yeast EB1 protein effectively substitutes for the human protein but also requires C-APC in promoting microtubule polymerization. Finally, C-APC is able to promote microtubule polymerization when stably expressed in APC mutant cells, demonstrating the ability of C-APC to promote microtubule assembly in vivo. Thus, the interaction between EB1 and APC plays an essential role in the regulation of microtubule polymerization, and a similar mechanism may be conserved in yeast.

AB - Human EB1 was originally cloned as a protein that interacts with the COOH terminus of adenomatous polyposis coli (APC) [1]. Interestingly, this interaction is often disrupted in colon cancer, due to mutations in APC. EB1 also interacts with the plus-ends of microtubules and targets APC to microtubule tips [2-6]. Since APC is detected on the kinetochores of chromosomes, it has been hypothesized that the EB1-APC interaction connects microtubule spindles to the kinetochores and regulates microtubule stability [7-9]. In yeast, EB1 regulates microtubule dynamics [6, 10, 11], and its binding domain in APC may be conserved in Kar9, an EB1 binding protein involved in the microtubule-capturing mechanism [12-14]. These results suggest that the interaction of EB1 and APC is important and may be conserved. However, it is largely unknown whether the EB1-APC interaction affects microtubule dynamics. Here, we show that EB1 potently promotes microtubule polymerization in vitro and in permeabilized cells, but, surprisingly, only in the presence of the COOH-terminal EB1 binding domain of APC (C-APC). Significantly, this C-APC activity is abolished by phosphorylation, which also disrupts its ability to bind to EB1. Furthermore, yeast EB1 protein effectively substitutes for the human protein but also requires C-APC in promoting microtubule polymerization. Finally, C-APC is able to promote microtubule polymerization when stably expressed in APC mutant cells, demonstrating the ability of C-APC to promote microtubule assembly in vivo. Thus, the interaction between EB1 and APC plays an essential role in the regulation of microtubule polymerization, and a similar mechanism may be conserved in yeast.

UR - http://www.scopus.com/inward/record.url?scp=0035838422&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035838422&partnerID=8YFLogxK

U2 - 10.1016/S0960-9822(01)00297-4

DO - 10.1016/S0960-9822(01)00297-4

M3 - Article

C2 - 11470413

AN - SCOPUS:0035838422

VL - 11

SP - 1062

EP - 1067

JO - Current Biology

JF - Current Biology

SN - 0960-9822

IS - 13

ER -