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Juxtamembrane localization of the protein phosphatase-1 inhibitor protein PHI-1 in smooth muscle cells

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Abstract

Protein phosphorylation regulates many fundamental processes and protein phosphatase-1 (PP1) is a major phosphatase that determines the levels of Ser/Thr phosphorylation. Regulatory subunits and inhibitor phosphoproteins control PP1 activity. PHI-1 is a member of a family of PP1 inhibitor phosphoproteins that was discovered based on sequence similarity to the known inhibitor CPI-17. To learn more about PHI-1 we determined the tissue distribution of PHI-1 in embryonic and adult tissues, and examined its cellular localization by immunohistochemistry. In the embryo PHI-1 appeared first in the heart at E10, and by E15 it was detected in multiple tissues. Expression in adult tissues was strikingly different, with PHI-1 detected primarily in smooth muscles in the intestine, blood vessels, and male and female genitourinary tracts. PHI-1 also was highly expressed in the endothelial layer of blood vessels. Both PHI-1 and CPI-17 are expressed predominantly in adult smooth muscles. Whereas CPI-17 staining was diffuse PHI-1 was concentrated along the cell membrane in distinct foci, detected by confocal and electron microscopy. The common tissue distribution but different cellular localization of PHI-1 and CPI-17 suggest distinctive physiological roles for these two PP1 inhibitors.

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Acknowledgements

We would like to thank Jan Redick of the University of Virginia Advanced Microscopy Facility for assistance with confocal and electron microscopy. This research was supported by grants (to D.L.B.) CA40042 from the National Cancer Institute and GM56362 from the National Institute of General Medical Sciences, US Public Health Service.

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Author notes

  1. Allen D. Everett

    Present address: Department of Pediatrics, Johns Hopkins School of Medicine, Baltimore, MD 21287, USA

Authors and Affiliations

  1. Center for Cell Signaling, University of Virginia School of Medicine, P.O. Box 800577, Charlottesville, VA 22908, USA

    Nikolaos A. Tountas & David L. Brautigan

  2. Department of Pathology (Neuropathology), University of Virginia Health System, Charlottesville, VA 22908, USA

    James W. Mandell

  3. Department of Pediatrics and the Cardiovascular Research Center, University of Virginia Health System, Charlottesville, VA 22908, USA

    Allen D. Everett

Authors
  1. Nikolaos A. Tountas
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  2. James W. Mandell
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  3. Allen D. Everett
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  4. David L. Brautigan
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Corresponding author

Correspondence to David L. Brautigan.

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Tountas, N.A., Mandell, J.W., Everett, A.D. et al. Juxtamembrane localization of the protein phosphatase-1 inhibitor protein PHI-1 in smooth muscle cells. Histochem Cell Biol 121, 343–350 (2004). https://doi.org/10.1007/s00418-004-0642-8

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  • DOI: https://doi.org/10.1007/s00418-004-0642-8

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