Tài liệu Báo cáo khoa học: Interaction between very-KIND Ras guanine exchange factor and

Interaction between very-KIND Ras guanine exchange

factor and microtubule-associated protein 2, and its role

in dendrite growth – structure and function of the second

kinase noncatalytic C-lobe domain

Jinhong Huang1,*, Asako Furuya1

, Kanehiro Hayashi1–3 and Teiichi Furuichi1,2,4

1 Laboratory for Molecular Neurogenesis, RIKEN Brain Science Institute, Saitama, Japan

2 JST, CREST, Kawaguchi, Saitama, Japan

3 Research Institute of Pharmaceutical Sciences, Musashino University, Tokyo, Japan

4 Faculty of Science and Technology, Tokyo University of Science, Chiba, Japan

Keywords

dendrite growth; KIND domain; MAP2;

protein–protein interaction; RasGEF

Correspondence

T. Furuichi, Laboratory for Molecular

Neurogenesis, RIKEN Brain Science

Institute, 2-1 Hirosawa, Wako 351-0198,

Japan

Fax: +81 48 467 6079

Tel: +81 48 467 5906

E-mail: [email protected]

*Present address

Discovery & Development Laboratory I,

Hanno Research Center, Taiho

Pharmaceutical Co., Ltd, Saitama, Japan

(Received 5 January 2011, revised 19

February 2011, accepted 28 February

2011)

doi:10.1111/j.1742-4658.2011.08085.x

The kinase noncatalytic C-lobe domain (KIND) is a putative protein–protein

interaction module. Four KIND-containing proteins, Spir-2 (actin-nuclear

factor), PTPN13 (protein tyrosine phosphatase), FRMPD2 (scaffold protein)

and very-KIND (v-KIND) (brain-specific Ras guanine nucleotide exchange

factor), have been identified to date. Uniquely, v-KIND has two KINDs (i.e.

KIND1 and KIND2), whereas the other three proteins have only one. The

functional role of KIND, however, remains unclear. We previously demon￾strated that v-KIND interacts with the high-molecular weight microtubule￾associated protein 2 (MAP2), a dendritic microtubule-associated protein,

leading to negative regulation of neuronal dendrite growth. In the present

study, we analyzed the structure–function relationships of the v-KIND–

MAP2 interaction by generating a series of mutant constructs. The interac￾tion with endogenous MAP2 in mouse cerebellar granule cells was specific to

v-KIND KIND2, but not KIND1, and was not observed for the KINDs

from other KIND-containing proteins. The binding core modules critical for

the v-KIND–MAP2 interaction were defined within 32 residues of the mouse

v-KIND KIND2 and 43 residues of the mouse MAP2 central domain. Three

Leu residues at amino acid positions 461, 474 and 477 in the MAP2-binding

core module of KIND2 contributed to the interaction. The MAP2-binding

core module itself promoted dendrite branching as a dominant-negative regu￾lator of v-KIND in hippocampal neurons. The results reported in the present

study demonstrate the structural and functional determinant underlying the

v-KIND–MAP2 interaction that controls dendrite arborization patterns.

Structured digital abstract

l vKIND-KIND2 binds to Map2 by pull down (View interaction)

l Map2 physically interacts with vKIND-KIND2 by pull down (View interaction 1, 2, 3, 4, 5)

l Map2 physically interacts with vKIND by pull down (View interaction)

l Map2 physically interacts with vKIND-KIND2 by anti bait coimmunoprecipitation (View interaction)

l vKIND-KIND2 physically interacts with Map2 by pull down (View interaction)

Abbreviations

CD, central domain; DIV, day in vitro; EGFP, enhanced green fluorescent protein; GST, glutathione S-transferase; HMW, high-molecular￾weight; KIND, kinase noncatalytic C-lobe domain; KIND1, first kinase noncatalytic C-lobe domain; KIND2, second kinase noncatalytic C-lobe

domain; MAP2, microtubule-associated protein 2; GEF, guanine exchange factor; v-KIND, very-KIND.

FEBS Journal 278 (2011) 1651–1661 ª 2011 The Authors Journal compilation ª 2011 FEBS 1651