Molecular and Cellular Biochemistry

, Volume 350, Issue 1–2, pp 201–206 | Cite as

Phosphorylation of Caldesmon by PFTAIRE1 kinase promotes actin binding and formation of stress fibers

  • Wilson K. C. Leung
  • Arthur K. K. Ching
  • Nathalie Wong
Article

Abstract

Caldesmon (CaD) is an actin-binding protein that is capable of stabilizing actin filaments. Phosphorylation of CaD is widely accepted in the actin cytoskeletal modeling and promotion of cell migration. In this study, we show that CaD is a downstream phosphorylation substrate of PFTK1, a novel Cdc-2-related ser/thr protein kinase. Our study stemmed from an earlier investigation where we demonstrated that PFTK1 kinase conferred cell migratory advantages in human hepatocellular carcinoma (HCC) cells. Here, we showed that PFTK1-knockdown cells exhibited much reduced CaD phosphorylation and consequently caused dissociation of CaD from the F-actin fibers. The cellular localization of CaD was also altered in the absence of PFTK1. Immunofluorescence analysis revealed that PFTK1-abrogated cells exhibited a diffused and blurred appearance of CaD localization, whereas intact co-localization with F-actins was apparent in PFTK1-expressing cells. Without the binding of CaD to actin, disappearance of actin stress fibers was also evident in PFTK1-abrogated cells. In addition, we found that CaD is also commonly up-regulated in HCC tumors when compared to adjacent non-malignant liver (P = 0.022). Taken together, our results highlight a novel biological cascade that involved the phosphorylation activation of CaD by PFTK1 kinase in promoting formation of actin stress fibers.

Keywords

PFTK1 Caldesmon Actin binding 

Abbreviations

PBS

Phosphate buffered saline

qRT-PCR

Quantitative reverse-transcriptase polymerase chain reaction

SDS-PAGE

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

TRITC

Tetramethylrhodamine B isothiocyanate

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Copyright information

© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  • Wilson K. C. Leung
    • 1
    • 2
  • Arthur K. K. Ching
    • 1
    • 2
  • Nathalie Wong
    • 1
    • 2
  1. 1.Department of Anatomical and Cellular Pathology at the Li Ka-Shing Institute of Health Sciences, Prince of Wales HospitalChinese University of Hong KongHong KongChina
  2. 2.State Key Laboratory in Oncology in South ChinaChinese University of Hong KongHong KongChina

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