PNUTS (phosphatase nuclear targeting subunit) inhibits retinoblastoma-directed PP1 activity.
Protein phosphatase type 1 catalytic subunit (PP1c) is a serine/threonine phosphatase involved in the dephosphorylation of many proteins in eukaryotic cells. It associates with several known targeting or regulatory subunits that directly regulate PP1c activity toward specific substrates. The recently identified Phosphatase Nuclear Targeting Subunit (PNUTS) binds to PP1c and inhibits PP1 activity toward phosphorylase a. One of the substrates of PP1c has been shown to be the cell cycle regulatory protein, Retinoblastoma (pRb). In this study, we show that PNUTS dissociates from PP1c under mildly hypoxic cell growth conditions that lead to an increase of PP1c activity toward pRb. We developed an assay that measures pRb-directed PP1c activity and show that a GST-PNUTS fusion protein inhibits phosphatase activity toward pRb when using PP1c from cell lysates, GST-PP1c, or purified PP1c. These studies suggest that PNUTS is involved in the regulation of PP1c activity toward pRb.
Udho
Eshwar
E
Department of Biological Sciences, Dyson College of Arts and Sciences, Pace University-Pleasantville, 861 Bedford Road, Marks Hall, Pleasantville, NY 10570-2799, USA.
Tedesco
Vivienne C
VC
Zygmunt
Adam
A
Krucher
Nancy A
NA
D000818:Animals; D002352:Carrier Proteins / Q000494:pharmacology*; D015687:Cell Hypoxia / Q000502:physiology*; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D047908:Intracellular Signaling Peptides and Proteins; D007668:Kidney; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors*; D054645:Protein Phosphatase 1; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000494:pharmacology; D016160:Retinoblastoma Protein / Q000187:drug effects / Q000378:metabolism*; D013379:Substrate Specificity; D014162:Transfection; D019459:Urothelium
D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D021122:Protein Subunits; D011994:Recombinant Proteins; D016160:Retinoblastoma Protein; C021042:protein phosphatase inhibitor-1; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1