Parkin interacts with LIM Kinase 1 and reduces its cofilin-phosphorylation activity via ubiquitination

Meng K. Lim, Takeshi Kawamura, Yosuke Ohsawa, Masafumi Ohtsubo, Shuichi Asakawa, Atsushi Takayanagi, Nobuyoshi Shimizu

Research output: Contribution to journalArticle

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Abstract

Mutations in the PARKIN (PARK2) gene have been found in the majority of early-onset familial Parkinson's disease (PD) patients with autosomal recessive juvenile parkinsonism (ARJP). Parkin protein functions as an ubiquitin (E3) ligase that targets specific proteins for degradation in the 26S proteasome. Here, based on a mass spectrometry analysis of the human dopaminergic neuroblastoma-derived cell line SH-SY5Y that over-expresses parkin, we found that parkin may suppress cofilin phosphorylation. LIM Kinase 1 (LIMK1) is the upstream protein that phosphorylates cofilin, an actin depolymerizing protein. Thus, we postulated a possible connection between parkin and LIMK1. Our studies in other cell lines, using co-transfection assays, demonstrated that LIMK1 and parkin bind each other. LIMK1 also interacted with previously known parkin interactors Hsp70 and CHIP. Parkin enhanced LIMK1-ubiquitination in the human neuroblastoma-derived BE(2)-M17 cell line, but not in the human embryonic kidney-derived HEK293 cell line. In fact, parkin-over-expression reduced the level of LIMK1-induced phosphocofilin in the BE(2)-M17 cells but not in the HEK293 cells. Additionally, in simian kidney-derived COS-7 cells, parkin-over-expression reduced LIMK1-induced actin filament accumulation. LIMK1 in cultured cells regulates parkin reversibly: LIMK1 did not phosphorylate parkin but LIMK1 overexpression reduced parkin self-ubiquitination in vitro and in HEK293 cells. Furthermore, in the cells co-transfected with parkin and p38, LIMK1 significantly decreased p38-ubiquitination by parkin. These findings demonstrate a cell-type dependent functional interaction between parkin and LIMK1 and provide new evidence that links parkin and LIMK1 in the pathogenesis of familial PD.

Original languageEnglish
Pages (from-to)2858-2874
Number of pages17
JournalExperimental Cell Research
Volume313
Issue number13
DOIs
Publication statusPublished - 2007 Aug 1

Fingerprint

Lim Kinases
Actin Depolymerizing Factors
Ubiquitination
Phosphorylation
HEK293 Cells
Cell Line
Neuroblastoma
Parkinson Disease
Kidney
Ubiquitin-Protein Ligases
COS Cells
Parkinsonian Disorders

Keywords

  • Autosomal recessive juvenile parkinsonism (ARJP)
  • Co-immunoprecipitation
  • Cofilin
  • LIM Kinase 1
  • PARKIN
  • Parkinson's disease
  • Phosphocofilin
  • Phosphorylation
  • Ubiquitination

ASJC Scopus subject areas

  • Cell Biology

Cite this

Lim, M. K., Kawamura, T., Ohsawa, Y., Ohtsubo, M., Asakawa, S., Takayanagi, A., & Shimizu, N. (2007). Parkin interacts with LIM Kinase 1 and reduces its cofilin-phosphorylation activity via ubiquitination. Experimental Cell Research, 313(13), 2858-2874. https://doi.org/10.1016/j.yexcr.2007.04.016

Parkin interacts with LIM Kinase 1 and reduces its cofilin-phosphorylation activity via ubiquitination. / Lim, Meng K.; Kawamura, Takeshi; Ohsawa, Yosuke; Ohtsubo, Masafumi; Asakawa, Shuichi; Takayanagi, Atsushi; Shimizu, Nobuyoshi.

In: Experimental Cell Research, Vol. 313, No. 13, 01.08.2007, p. 2858-2874.

Research output: Contribution to journalArticle

Lim, MK, Kawamura, T, Ohsawa, Y, Ohtsubo, M, Asakawa, S, Takayanagi, A & Shimizu, N 2007, 'Parkin interacts with LIM Kinase 1 and reduces its cofilin-phosphorylation activity via ubiquitination', Experimental Cell Research, vol. 313, no. 13, pp. 2858-2874. https://doi.org/10.1016/j.yexcr.2007.04.016
Lim, Meng K. ; Kawamura, Takeshi ; Ohsawa, Yosuke ; Ohtsubo, Masafumi ; Asakawa, Shuichi ; Takayanagi, Atsushi ; Shimizu, Nobuyoshi. / Parkin interacts with LIM Kinase 1 and reduces its cofilin-phosphorylation activity via ubiquitination. In: Experimental Cell Research. 2007 ; Vol. 313, No. 13. pp. 2858-2874.
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abstract = "Mutations in the PARKIN (PARK2) gene have been found in the majority of early-onset familial Parkinson's disease (PD) patients with autosomal recessive juvenile parkinsonism (ARJP). Parkin protein functions as an ubiquitin (E3) ligase that targets specific proteins for degradation in the 26S proteasome. Here, based on a mass spectrometry analysis of the human dopaminergic neuroblastoma-derived cell line SH-SY5Y that over-expresses parkin, we found that parkin may suppress cofilin phosphorylation. LIM Kinase 1 (LIMK1) is the upstream protein that phosphorylates cofilin, an actin depolymerizing protein. Thus, we postulated a possible connection between parkin and LIMK1. Our studies in other cell lines, using co-transfection assays, demonstrated that LIMK1 and parkin bind each other. LIMK1 also interacted with previously known parkin interactors Hsp70 and CHIP. Parkin enhanced LIMK1-ubiquitination in the human neuroblastoma-derived BE(2)-M17 cell line, but not in the human embryonic kidney-derived HEK293 cell line. In fact, parkin-over-expression reduced the level of LIMK1-induced phosphocofilin in the BE(2)-M17 cells but not in the HEK293 cells. Additionally, in simian kidney-derived COS-7 cells, parkin-over-expression reduced LIMK1-induced actin filament accumulation. LIMK1 in cultured cells regulates parkin reversibly: LIMK1 did not phosphorylate parkin but LIMK1 overexpression reduced parkin self-ubiquitination in vitro and in HEK293 cells. Furthermore, in the cells co-transfected with parkin and p38, LIMK1 significantly decreased p38-ubiquitination by parkin. These findings demonstrate a cell-type dependent functional interaction between parkin and LIMK1 and provide new evidence that links parkin and LIMK1 in the pathogenesis of familial PD.",
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AB - Mutations in the PARKIN (PARK2) gene have been found in the majority of early-onset familial Parkinson's disease (PD) patients with autosomal recessive juvenile parkinsonism (ARJP). Parkin protein functions as an ubiquitin (E3) ligase that targets specific proteins for degradation in the 26S proteasome. Here, based on a mass spectrometry analysis of the human dopaminergic neuroblastoma-derived cell line SH-SY5Y that over-expresses parkin, we found that parkin may suppress cofilin phosphorylation. LIM Kinase 1 (LIMK1) is the upstream protein that phosphorylates cofilin, an actin depolymerizing protein. Thus, we postulated a possible connection between parkin and LIMK1. Our studies in other cell lines, using co-transfection assays, demonstrated that LIMK1 and parkin bind each other. LIMK1 also interacted with previously known parkin interactors Hsp70 and CHIP. Parkin enhanced LIMK1-ubiquitination in the human neuroblastoma-derived BE(2)-M17 cell line, but not in the human embryonic kidney-derived HEK293 cell line. In fact, parkin-over-expression reduced the level of LIMK1-induced phosphocofilin in the BE(2)-M17 cells but not in the HEK293 cells. Additionally, in simian kidney-derived COS-7 cells, parkin-over-expression reduced LIMK1-induced actin filament accumulation. LIMK1 in cultured cells regulates parkin reversibly: LIMK1 did not phosphorylate parkin but LIMK1 overexpression reduced parkin self-ubiquitination in vitro and in HEK293 cells. Furthermore, in the cells co-transfected with parkin and p38, LIMK1 significantly decreased p38-ubiquitination by parkin. These findings demonstrate a cell-type dependent functional interaction between parkin and LIMK1 and provide new evidence that links parkin and LIMK1 in the pathogenesis of familial PD.

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