PKN binds and phosphorylates human papillomavirus E6 oncoprotein

Qingshen Gao, Ajay Kumar, Seetha Srinivasan, Latika Singh, Hideyuki Mukai, Yoshitaka Ono, David E. Wazer, Vimla Band

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

The high risk human papillomaviruses (HPVs) are associated with carcinomas of cervix and other genital tumors. Previous studies have identified two viral oncoproteins E6 and E7, which are expressed in the majority of HPV-associated carcinomas. The ability of high risk HPV E6 protein to immortalize human mammary epithelial cells has provided a single gene model to study the mechanisms of E6-induced oncogenic transformation. In recent years, it has become clear that in addition to E6-induced degradation of p53 tumor suppressor protein, other targets of E6 are required for mammary epithelial cells immortalization. Using the yeast two-hybrid system, we have identified a novel interaction of HPV16 E6 with protein kinase PKN, a fatty acid- and Rho small G protein-activated serine/threonine kinase with a catalytic domain highly homologous to protein kinase C. We demonstrate direct binding of high risk HPV E6 proteins to PKN in wheat-germ lysate in vitro and in 293T cells in vivo. Importantly, E6 proteins of high risk HPVs but not low risk HPVs were able to bind PKN. Furthermore, all the immortalization- competent and many immortalization-non-competent E6 mutants bind PKN. These data suggest that binding to PKN may be required but not sufficient for immortalizing normal mammary epithelial cells. Finally, we show that PKN phosphorylates E6, demonstrating for the first time that HPV E6 is a phosphoprotein. Our finding suggests a novel link between HPV E6 mediated oncogenesis and regulation of a well known phosphorylation cascade.

Original languageEnglish (US)
Pages (from-to)14824-14830
Number of pages7
JournalJournal of Biological Chemistry
Volume275
Issue number20
DOIs
StatePublished - May 19 2000

Fingerprint

Oncogene Proteins
Breast
Tumor Suppressor Protein p53
Epithelial Cells
Phosphorylation
Proteins
Monomeric GTP-Binding Proteins
Protein-Serine-Threonine Kinases
Phosphoproteins
Hybrid systems
Yeast
Protein Kinases
Protein Kinase C
Tumors
Fatty Acids
Genes
Carcinoma
rho GTP-Binding Proteins
Two-Hybrid System Techniques
Degradation

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Gao, Q., Kumar, A., Srinivasan, S., Singh, L., Mukai, H., Ono, Y., ... Band, V. (2000). PKN binds and phosphorylates human papillomavirus E6 oncoprotein. Journal of Biological Chemistry, 275(20), 14824-14830. https://doi.org/10.1074/jbc.275.20.14824

PKN binds and phosphorylates human papillomavirus E6 oncoprotein. / Gao, Qingshen; Kumar, Ajay; Srinivasan, Seetha; Singh, Latika; Mukai, Hideyuki; Ono, Yoshitaka; Wazer, David E.; Band, Vimla.

In: Journal of Biological Chemistry, Vol. 275, No. 20, 19.05.2000, p. 14824-14830.

Research output: Contribution to journalArticle

Gao, Q, Kumar, A, Srinivasan, S, Singh, L, Mukai, H, Ono, Y, Wazer, DE & Band, V 2000, 'PKN binds and phosphorylates human papillomavirus E6 oncoprotein', Journal of Biological Chemistry, vol. 275, no. 20, pp. 14824-14830. https://doi.org/10.1074/jbc.275.20.14824
Gao Q, Kumar A, Srinivasan S, Singh L, Mukai H, Ono Y et al. PKN binds and phosphorylates human papillomavirus E6 oncoprotein. Journal of Biological Chemistry. 2000 May 19;275(20):14824-14830. https://doi.org/10.1074/jbc.275.20.14824
Gao, Qingshen ; Kumar, Ajay ; Srinivasan, Seetha ; Singh, Latika ; Mukai, Hideyuki ; Ono, Yoshitaka ; Wazer, David E. ; Band, Vimla. / PKN binds and phosphorylates human papillomavirus E6 oncoprotein. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 20. pp. 14824-14830.
@article{6ff5363112344681b6e7adbbf3dd784c,
title = "PKN binds and phosphorylates human papillomavirus E6 oncoprotein",
abstract = "The high risk human papillomaviruses (HPVs) are associated with carcinomas of cervix and other genital tumors. Previous studies have identified two viral oncoproteins E6 and E7, which are expressed in the majority of HPV-associated carcinomas. The ability of high risk HPV E6 protein to immortalize human mammary epithelial cells has provided a single gene model to study the mechanisms of E6-induced oncogenic transformation. In recent years, it has become clear that in addition to E6-induced degradation of p53 tumor suppressor protein, other targets of E6 are required for mammary epithelial cells immortalization. Using the yeast two-hybrid system, we have identified a novel interaction of HPV16 E6 with protein kinase PKN, a fatty acid- and Rho small G protein-activated serine/threonine kinase with a catalytic domain highly homologous to protein kinase C. We demonstrate direct binding of high risk HPV E6 proteins to PKN in wheat-germ lysate in vitro and in 293T cells in vivo. Importantly, E6 proteins of high risk HPVs but not low risk HPVs were able to bind PKN. Furthermore, all the immortalization- competent and many immortalization-non-competent E6 mutants bind PKN. These data suggest that binding to PKN may be required but not sufficient for immortalizing normal mammary epithelial cells. Finally, we show that PKN phosphorylates E6, demonstrating for the first time that HPV E6 is a phosphoprotein. Our finding suggests a novel link between HPV E6 mediated oncogenesis and regulation of a well known phosphorylation cascade.",
author = "Qingshen Gao and Ajay Kumar and Seetha Srinivasan and Latika Singh and Hideyuki Mukai and Yoshitaka Ono and Wazer, {David E.} and Vimla Band",
year = "2000",
month = "5",
day = "19",
doi = "10.1074/jbc.275.20.14824",
language = "English (US)",
volume = "275",
pages = "14824--14830",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "20",

}

TY - JOUR

T1 - PKN binds and phosphorylates human papillomavirus E6 oncoprotein

AU - Gao, Qingshen

AU - Kumar, Ajay

AU - Srinivasan, Seetha

AU - Singh, Latika

AU - Mukai, Hideyuki

AU - Ono, Yoshitaka

AU - Wazer, David E.

AU - Band, Vimla

PY - 2000/5/19

Y1 - 2000/5/19

N2 - The high risk human papillomaviruses (HPVs) are associated with carcinomas of cervix and other genital tumors. Previous studies have identified two viral oncoproteins E6 and E7, which are expressed in the majority of HPV-associated carcinomas. The ability of high risk HPV E6 protein to immortalize human mammary epithelial cells has provided a single gene model to study the mechanisms of E6-induced oncogenic transformation. In recent years, it has become clear that in addition to E6-induced degradation of p53 tumor suppressor protein, other targets of E6 are required for mammary epithelial cells immortalization. Using the yeast two-hybrid system, we have identified a novel interaction of HPV16 E6 with protein kinase PKN, a fatty acid- and Rho small G protein-activated serine/threonine kinase with a catalytic domain highly homologous to protein kinase C. We demonstrate direct binding of high risk HPV E6 proteins to PKN in wheat-germ lysate in vitro and in 293T cells in vivo. Importantly, E6 proteins of high risk HPVs but not low risk HPVs were able to bind PKN. Furthermore, all the immortalization- competent and many immortalization-non-competent E6 mutants bind PKN. These data suggest that binding to PKN may be required but not sufficient for immortalizing normal mammary epithelial cells. Finally, we show that PKN phosphorylates E6, demonstrating for the first time that HPV E6 is a phosphoprotein. Our finding suggests a novel link between HPV E6 mediated oncogenesis and regulation of a well known phosphorylation cascade.

AB - The high risk human papillomaviruses (HPVs) are associated with carcinomas of cervix and other genital tumors. Previous studies have identified two viral oncoproteins E6 and E7, which are expressed in the majority of HPV-associated carcinomas. The ability of high risk HPV E6 protein to immortalize human mammary epithelial cells has provided a single gene model to study the mechanisms of E6-induced oncogenic transformation. In recent years, it has become clear that in addition to E6-induced degradation of p53 tumor suppressor protein, other targets of E6 are required for mammary epithelial cells immortalization. Using the yeast two-hybrid system, we have identified a novel interaction of HPV16 E6 with protein kinase PKN, a fatty acid- and Rho small G protein-activated serine/threonine kinase with a catalytic domain highly homologous to protein kinase C. We demonstrate direct binding of high risk HPV E6 proteins to PKN in wheat-germ lysate in vitro and in 293T cells in vivo. Importantly, E6 proteins of high risk HPVs but not low risk HPVs were able to bind PKN. Furthermore, all the immortalization- competent and many immortalization-non-competent E6 mutants bind PKN. These data suggest that binding to PKN may be required but not sufficient for immortalizing normal mammary epithelial cells. Finally, we show that PKN phosphorylates E6, demonstrating for the first time that HPV E6 is a phosphoprotein. Our finding suggests a novel link between HPV E6 mediated oncogenesis and regulation of a well known phosphorylation cascade.

UR - http://www.scopus.com/inward/record.url?scp=0034685758&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034685758&partnerID=8YFLogxK

U2 - 10.1074/jbc.275.20.14824

DO - 10.1074/jbc.275.20.14824

M3 - Article

C2 - 10809724

AN - SCOPUS:0034685758

VL - 275

SP - 14824

EP - 14830

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 20

ER -