The D Domain of LRRC4 anchors ERK1/2 in the cytoplasm and competitively inhibits MEK/ERK activation in glioma cells

J Hematol Oncol. 2016 Nov 25;9(1):130. doi: 10.1186/s13045-016-0355-1.

Authors

Zeyou Wang^{1

2

3

4}, Qin Guo³, Rong Wang^{1

2

3}, Gang Xu^{1

2

5}, Peiyao Li^{1

2

3}, Yingnan Sun⁶, Xiaoling She^{1

2

3

7}, Qiang Liu³, Qiong Chen⁶, Zhibin Yu^{1

2

3}, Changhong Liu^{1

2

3}, Jing Xiong^{1

2

8}, Guiyuan Li^{9

10

11}, Minghua Wu^{12

13

14}

Affiliations

¹ The Key Laboratory of Carcinogenesis of the Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, China.
² Cancer Research Institute, The Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Central South University, Changsha, Hunan, 410008, China.
³ Hunan Key Laboratory of Nonresolving Inflammation and Cancer, Disease Genome Research Center, The Third Xiangya Hospital, Central South University, Changsha, Hunan, 410013, China.
⁴ Department of Laboratory Medicine, The Second Xiangya Hospital, Central South University, Changsha, Hunan, 410011, China.
⁵ Medical College, University of South China, Hengyang, Hunan, 421001, China.
⁶ Hunan Provincial Tumor Hospital and the Affiliated Tumor Hospital of Xiangya Medical School, Central South University, Changsha, 410013, Hunan, China.
⁷ Department of Pathology, The Second Xiangya Hospital, Central South University, Changsha, Hunan, 410011, China.
⁸ Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, China.
⁹ The Key Laboratory of Carcinogenesis of the Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, China. lgy@csu.edu.cn.
¹⁰ Cancer Research Institute, The Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Central South University, Changsha, Hunan, 410008, China. lgy@csu.edu.cn.
¹¹ Hunan Key Laboratory of Nonresolving Inflammation and Cancer, Disease Genome Research Center, The Third Xiangya Hospital, Central South University, Changsha, Hunan, 410013, China. lgy@csu.edu.cn.
¹² The Key Laboratory of Carcinogenesis of the Chinese Ministry of Health, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, China. wuminghua554@aliyun.com.
¹³ Cancer Research Institute, The Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Central South University, Changsha, Hunan, 410008, China. wuminghua554@aliyun.com.
¹⁴ Hunan Key Laboratory of Nonresolving Inflammation and Cancer, Disease Genome Research Center, The Third Xiangya Hospital, Central South University, Changsha, Hunan, 410013, China. wuminghua554@aliyun.com.

Abstract

Background: As a well-characterized key player in various signal transduction networks, extracellular-signal-regulated kinase (ERK1/2) has been widely implicated in the development of many malignancies. We previously found that Leucine-rich repeat containing 4 (LRRC4) was a tumor suppressor and a negative regulator of the ERK/MAPK pathway in glioma tumorigenesis. However, the precise molecular role of LRRC4 in ERK signal transmission is unclear.

Methods: The interaction between LRRC4 and ERK1/2 was assessed by co-immunoprecipitation and GST pull-down assays in vivo and in vitro. We also investigated the interaction of LRRC4 and ERK1/2 and the role of the D domain in ERK activation in glioma cells.

Results: Here, we showed that LRRC4 and ERK1/2 interact via the D domain and CD domain, respectively. Following EGF stimuli, the D domain of LRRC4 anchors ERK1/2 in the cytoplasm and abrogates ERK1/2 activation and nuclear translocation. In glioblastoma cells, ectopic LRRC4 expression competitively inhibited the interaction of endogenous mitogen-activated protein kinase (MEK) and ERK1/2. Mutation of the D domain decreased the LRRC4-mediated inhibition of MAPK signaling and its anti-proliferation and anti-invasion roles.

Conclusions: Our results demonstrated that the D domain of LRRC4 anchors ERK1/2 in the cytoplasm and competitively inhibits MEK/ERK activation in glioma cells. These findings identify a new mechanism underlying glioblastoma progression and suggest a novel therapeutic strategy by restoring the activity of LRRC4 to decrease MAPK cascade activation.

Keywords: CD domain; D domain; ERK1/2; Leucine-rich repeat; MAPK.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cytoplasm / enzymology
Cytoplasm / metabolism
Disease Progression
Enzyme Activation
Glioma / enzymology*
Glioma / pathology
Humans
MAP Kinase Signaling System / drug effects*
Nerve Tissue Proteins / chemistry*
Nerve Tissue Proteins / pharmacology
Protein Domains / physiology*
Protein Kinase Inhibitors / pharmacology

Substances

LRRC4 protein, human
Nerve Tissue Proteins
Protein Kinase Inhibitors