Mitotic phosphorylation activates hepatoma-derived growth factor as a mitogen

Allen D Everett, Jun Yang, Monzur Rahman, Pratima Dulloor, David L. Brautigan

Research output: Contribution to journalArticle

Abstract

Background: Hepatoma-derived growth factor (HDGF) is a nuclear protein that is a mitogen for a wide variety of cells. Mass spectrometry based methods have identified HDGF as a phosphoprotein without validation or a functional consequence of this post-translational modification.Results: We found that HDGF in primary mouse aortic vascular smooth muscle cells (VSMC) was phosphorylated. Wild type HDGF was phosphorylated in asynchronous cells and substitution of S103, S165 and S202 to alanine each demonstrated a decrease in HDGF phosphorylation. A phospho-S103 HDGF specific antibody was developed and demonstrated mitosis-specific phosphorylation. HDGF-S103A was not mitogenic and FACS analysis demonstrated a G2/M arrest in HDGF-S103A expressing cells, whereas cells expressing HDGF-S103D showed cell cycle progression. Nocodazole arrest increased S103 phosphorylation from 1.6% to 29% (P = 0.037).Conclusions: Thus, HDGF is a phosphoprotein and phosphorylation of S103 is mitosis related and required for its function as a mitogen. We speculate that cell cycle regulated phosphorylation of HDGF may play an important role in vascular cell proliferation.

Original languageEnglish (US)
Article number15
JournalBMC Cell Biology
Volume12
DOIs
StatePublished - Apr 13 2011

Fingerprint

Mitogens
Phosphorylation
Phosphoproteins
Mitosis
Cell Cycle
hepatoma-derived growth factor
Nocodazole
Post Translational Protein Processing
Nuclear Proteins
Vascular Smooth Muscle
Alanine
Smooth Muscle Myocytes
Blood Vessels
Mass Spectrometry
Cell Proliferation
Antibodies

ASJC Scopus subject areas

  • Cell Biology

Cite this

Mitotic phosphorylation activates hepatoma-derived growth factor as a mitogen. / Everett, Allen D; Yang, Jun; Rahman, Monzur; Dulloor, Pratima; Brautigan, David L.

In: BMC Cell Biology, Vol. 12, 15, 13.04.2011.

Research output: Contribution to journalArticle

Everett, Allen D ; Yang, Jun ; Rahman, Monzur ; Dulloor, Pratima ; Brautigan, David L. / Mitotic phosphorylation activates hepatoma-derived growth factor as a mitogen. In: BMC Cell Biology. 2011 ; Vol. 12.
@article{07ff3033f86e42dc8b382d172319a9d9,
title = "Mitotic phosphorylation activates hepatoma-derived growth factor as a mitogen",
abstract = "Background: Hepatoma-derived growth factor (HDGF) is a nuclear protein that is a mitogen for a wide variety of cells. Mass spectrometry based methods have identified HDGF as a phosphoprotein without validation or a functional consequence of this post-translational modification.Results: We found that HDGF in primary mouse aortic vascular smooth muscle cells (VSMC) was phosphorylated. Wild type HDGF was phosphorylated in asynchronous cells and substitution of S103, S165 and S202 to alanine each demonstrated a decrease in HDGF phosphorylation. A phospho-S103 HDGF specific antibody was developed and demonstrated mitosis-specific phosphorylation. HDGF-S103A was not mitogenic and FACS analysis demonstrated a G2/M arrest in HDGF-S103A expressing cells, whereas cells expressing HDGF-S103D showed cell cycle progression. Nocodazole arrest increased S103 phosphorylation from 1.6{\%} to 29{\%} (P = 0.037).Conclusions: Thus, HDGF is a phosphoprotein and phosphorylation of S103 is mitosis related and required for its function as a mitogen. We speculate that cell cycle regulated phosphorylation of HDGF may play an important role in vascular cell proliferation.",
author = "Everett, {Allen D} and Jun Yang and Monzur Rahman and Pratima Dulloor and Brautigan, {David L.}",
year = "2011",
month = "4",
day = "13",
doi = "10.1186/1471-2121-12-15",
language = "English (US)",
volume = "12",
journal = "BMC Cell Biology",
issn = "1471-2121",
publisher = "BioMed Central",

}

TY - JOUR

T1 - Mitotic phosphorylation activates hepatoma-derived growth factor as a mitogen

AU - Everett, Allen D

AU - Yang, Jun

AU - Rahman, Monzur

AU - Dulloor, Pratima

AU - Brautigan, David L.

PY - 2011/4/13

Y1 - 2011/4/13

N2 - Background: Hepatoma-derived growth factor (HDGF) is a nuclear protein that is a mitogen for a wide variety of cells. Mass spectrometry based methods have identified HDGF as a phosphoprotein without validation or a functional consequence of this post-translational modification.Results: We found that HDGF in primary mouse aortic vascular smooth muscle cells (VSMC) was phosphorylated. Wild type HDGF was phosphorylated in asynchronous cells and substitution of S103, S165 and S202 to alanine each demonstrated a decrease in HDGF phosphorylation. A phospho-S103 HDGF specific antibody was developed and demonstrated mitosis-specific phosphorylation. HDGF-S103A was not mitogenic and FACS analysis demonstrated a G2/M arrest in HDGF-S103A expressing cells, whereas cells expressing HDGF-S103D showed cell cycle progression. Nocodazole arrest increased S103 phosphorylation from 1.6% to 29% (P = 0.037).Conclusions: Thus, HDGF is a phosphoprotein and phosphorylation of S103 is mitosis related and required for its function as a mitogen. We speculate that cell cycle regulated phosphorylation of HDGF may play an important role in vascular cell proliferation.

AB - Background: Hepatoma-derived growth factor (HDGF) is a nuclear protein that is a mitogen for a wide variety of cells. Mass spectrometry based methods have identified HDGF as a phosphoprotein without validation or a functional consequence of this post-translational modification.Results: We found that HDGF in primary mouse aortic vascular smooth muscle cells (VSMC) was phosphorylated. Wild type HDGF was phosphorylated in asynchronous cells and substitution of S103, S165 and S202 to alanine each demonstrated a decrease in HDGF phosphorylation. A phospho-S103 HDGF specific antibody was developed and demonstrated mitosis-specific phosphorylation. HDGF-S103A was not mitogenic and FACS analysis demonstrated a G2/M arrest in HDGF-S103A expressing cells, whereas cells expressing HDGF-S103D showed cell cycle progression. Nocodazole arrest increased S103 phosphorylation from 1.6% to 29% (P = 0.037).Conclusions: Thus, HDGF is a phosphoprotein and phosphorylation of S103 is mitosis related and required for its function as a mitogen. We speculate that cell cycle regulated phosphorylation of HDGF may play an important role in vascular cell proliferation.

UR - http://www.scopus.com/inward/record.url?scp=79953880876&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79953880876&partnerID=8YFLogxK

U2 - 10.1186/1471-2121-12-15

DO - 10.1186/1471-2121-12-15

M3 - Article

C2 - 21489262

AN - SCOPUS:79953880876

VL - 12

JO - BMC Cell Biology

JF - BMC Cell Biology

SN - 1471-2121

M1 - 15

ER -