BAD, a Proapoptotic Protein, Escapes ERK/RSK Phosphorylation in Deguelin and siRNA-Treated HeLa Cells

Samra Hafeez; Mahwish Urooj; Shamiala Saleem; Zeeshan Gillani; Sumaira Shaheen; Mahmood Husain Qazi; Muhammad Imran Naseer; Zafar Iqbal; Shakeel Ahmed Ansari; Absarul Haque; Muhammad Asif; Manzoor Ahmad Mir; Ashraf Ali; Peter Natesan Pushparaj; Mohammad Sarwar Jamal; Mahmood Rasool

doi:10.1371/journal.pone.0145780

BAD, a Proapoptotic Protein, Escapes ERK/RSK Phosphorylation in Deguelin and siRNA-Treated HeLa Cells

PLoS One. 2016 Jan 8;11(1):e0145780. doi: 10.1371/journal.pone.0145780. eCollection 2016.

Authors

Samra Hafeez¹, Mahwish Urooj¹, Shamiala Saleem¹, Zeeshan Gillani¹, Sumaira Shaheen¹, Mahmood Husain Qazi¹, Muhammad Imran Naseer², Zafar Iqbal³, Shakeel Ahmed Ansari², Absarul Haque⁴, Muhammad Asif⁵, Manzoor Ahmad Mir⁶, Ashraf Ali⁴, Peter Natesan Pushparaj², Mohammad Sarwar Jamal⁴, Mahmood Rasool²

Affiliations

¹ Institute of Molecular Biology and Biotechnology and Center for Research in Molecular Medicine, the University of Lahore, Lahore, Pakistan.
² Center of Excellence in Genomic Medicine Research, King Abdulaziz University, Jeddah, Saudi Arabia.
³ College of Applied Medical Sciences, King Saud Bin Abdulaziz University for Health Sciences, National Guards Health Affairs, Riyadh, Saudi Arabia.
⁴ King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia.
⁵ Department of Biotechnology, BUITEMS, Quetta, Pakistan.
⁶ College of Applied Medical Science, Al Majmaah University, Majmaah City, Saudi Arabia.

Abstract

This study has been undertaken to explore the therapeutic effects of deguelin and specific siRNAs in HeLa cells. The data provided clearly show the silencing of ERK 1/2 with siRNAs and inhibition of ERK1/2 with deguelin treatment in HeLa cells. Additionally, we are providing information that deguelin binds directly to anti-apoptotic Bcl-2, Bcl-xl and Mcl-1 in the hydrophobic grooves, thereby releasing BAD and BAX from dimerization with these proteins. This results in increased apoptotic activity through the intrinsic pathway involved in rupture of mitochondrial membrane and release of cytochrome C. Evidence for inhibition of ERK1/2 by deguelin and escape of BAD phosphorylation at serine 112 through ERK/RSK pathway has been further fortified by obtaining similar results by silencing ERK 1/2 each with specific siRNAs. Increase in BAD after treatment with deguelin or siRNAs has been interpreted to mean that deguelin acts through several alternative pathways and therefore can be used as effective therapeutic agent.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Apoptosis / drug effects*
Binding Sites
Cytochromes c / metabolism
HeLa Cells
Humans
Mitochondrial Membranes / metabolism
Mitogen-Activated Protein Kinase 1 / antagonists & inhibitors
Mitogen-Activated Protein Kinase 1 / genetics
Mitogen-Activated Protein Kinase 1 / metabolism*
Mitogen-Activated Protein Kinase 3 / antagonists & inhibitors
Mitogen-Activated Protein Kinase 3 / genetics
Mitogen-Activated Protein Kinase 3 / metabolism*
Molecular Docking Simulation
Myeloid Cell Leukemia Sequence 1 Protein / chemistry
Myeloid Cell Leukemia Sequence 1 Protein / metabolism
Phosphorylation / drug effects
Protein Binding
Protein Structure, Tertiary
Proto-Oncogene Proteins c-bcl-2 / chemistry
Proto-Oncogene Proteins c-bcl-2 / metabolism
RNA Interference
Ribosomal Protein S6 Kinases / metabolism*
Rotenone / analogs & derivatives*
Rotenone / chemistry
Rotenone / pharmacology
bcl-Associated Death Protein / chemistry
bcl-Associated Death Protein / metabolism*
bcl-X Protein / chemistry
bcl-X Protein / metabolism

Substances

MCL1 protein, human
Myeloid Cell Leukemia Sequence 1 Protein
Proto-Oncogene Proteins c-bcl-2
bcl-Associated Death Protein
bcl-X Protein
Rotenone
Cytochromes c
Ribosomal Protein S6 Kinases
Mitogen-Activated Protein Kinase 1
Mitogen-Activated Protein Kinase 3
deguelin

Grants and funding

This work was funded by the National Plan for Science, Technology and Innovation (MAARIFAH)—King Abdulaziz City for Science and Technology—the Kingdom of Saudi Arabia—award number 12-BIO2267-03. The authors also acknowledge with thanks the Science and Technology Unit (STU), King Abdulaziz University for their excellent technical support.