Cargando…

hnRNPA1/UP1 Unfolds KRAS G-Quadruplexes and Feeds a Regulatory Axis Controlling Gene Expression

[Image: see text] Recent studies have proven that the genetic landscape of pancreatic cancer is dominated by the KRAS oncogene. Its transcription is controlled by a G-rich motif (called 32R) located immediately upstream of the TSS. 32R may fold into a G-quadruplex (G4) in equilibrium between two G4...

Descripción completa

Detalles Bibliográficos
Autores principales: Ferino, Annalisa, Marquevielle, Julien, Choudhary, Himanshi, Cinque, Giorgio, Robert, Coralie, Bourdoncle, Anne, Picco, Raffaella, Mergny, Jean-Louis, Salgado, Gilmar F., Xodo, Luigi E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2021
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8675163/
https://www.ncbi.nlm.nih.gov/pubmed/34926957
http://dx.doi.org/10.1021/acsomega.1c05538
_version_ 1784615824358637568
author Ferino, Annalisa
Marquevielle, Julien
Choudhary, Himanshi
Cinque, Giorgio
Robert, Coralie
Bourdoncle, Anne
Picco, Raffaella
Mergny, Jean-Louis
Salgado, Gilmar F.
Xodo, Luigi E.
author_facet Ferino, Annalisa
Marquevielle, Julien
Choudhary, Himanshi
Cinque, Giorgio
Robert, Coralie
Bourdoncle, Anne
Picco, Raffaella
Mergny, Jean-Louis
Salgado, Gilmar F.
Xodo, Luigi E.
author_sort Ferino, Annalisa
collection PubMed
description [Image: see text] Recent studies have proven that the genetic landscape of pancreatic cancer is dominated by the KRAS oncogene. Its transcription is controlled by a G-rich motif (called 32R) located immediately upstream of the TSS. 32R may fold into a G-quadruplex (G4) in equilibrium between two G4 conformers: G9T (T(M) = 61.2 °C) and G25T (T(M) = 54.7 °C). We found that both G4s bind to hnRNPA1 and its proteolytic fragment UP1, promoting several contacts with the RRM protein domains. 1D NMR analysis of DNA imino protons shows that, upon binding to UP1, G25T is readily unfolded at both 5′ and 3′ tetrads, while G9T is only partially unfolded. The impact of hnRNPA1 on KRAS expression was determined by comparing Panc-1 cells with two Panc-1 knockout cell lines in which hnRNPA1 was deleted by the CRISPR/Cas9 technology. The results showed that the expression of KRAS is inhibited in the knockout cell lines, indicating that hnRNPA1 is essential for the transcription of KRAS. In addition, the knockout cell lines, compared to normal Panc-1 cells, show a dramatic decrease in cell growth and capacity of colony formation. Pull-down and Western blot experiments indicate that conformer G25T is a better platform than conformer G9T for the assembly of the transcription preinitiation complex with PARP1, Ku70, MAZ, and hnRNPA1. Together, our data prove that hnRNPA1, being a key transcription factor for the activation of KRAS, can be a new therapeutic target for the rational design of anticancer strategies.
format Online
Article
Text
id pubmed-8675163
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher American Chemical Society
record_format MEDLINE/PubMed
spelling pubmed-86751632021-12-17 hnRNPA1/UP1 Unfolds KRAS G-Quadruplexes and Feeds a Regulatory Axis Controlling Gene Expression Ferino, Annalisa Marquevielle, Julien Choudhary, Himanshi Cinque, Giorgio Robert, Coralie Bourdoncle, Anne Picco, Raffaella Mergny, Jean-Louis Salgado, Gilmar F. Xodo, Luigi E. ACS Omega [Image: see text] Recent studies have proven that the genetic landscape of pancreatic cancer is dominated by the KRAS oncogene. Its transcription is controlled by a G-rich motif (called 32R) located immediately upstream of the TSS. 32R may fold into a G-quadruplex (G4) in equilibrium between two G4 conformers: G9T (T(M) = 61.2 °C) and G25T (T(M) = 54.7 °C). We found that both G4s bind to hnRNPA1 and its proteolytic fragment UP1, promoting several contacts with the RRM protein domains. 1D NMR analysis of DNA imino protons shows that, upon binding to UP1, G25T is readily unfolded at both 5′ and 3′ tetrads, while G9T is only partially unfolded. The impact of hnRNPA1 on KRAS expression was determined by comparing Panc-1 cells with two Panc-1 knockout cell lines in which hnRNPA1 was deleted by the CRISPR/Cas9 technology. The results showed that the expression of KRAS is inhibited in the knockout cell lines, indicating that hnRNPA1 is essential for the transcription of KRAS. In addition, the knockout cell lines, compared to normal Panc-1 cells, show a dramatic decrease in cell growth and capacity of colony formation. Pull-down and Western blot experiments indicate that conformer G25T is a better platform than conformer G9T for the assembly of the transcription preinitiation complex with PARP1, Ku70, MAZ, and hnRNPA1. Together, our data prove that hnRNPA1, being a key transcription factor for the activation of KRAS, can be a new therapeutic target for the rational design of anticancer strategies. American Chemical Society 2021-11-30 /pmc/articles/PMC8675163/ /pubmed/34926957 http://dx.doi.org/10.1021/acsomega.1c05538 Text en © 2021 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by-nc-nd/4.0/Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Ferino, Annalisa
Marquevielle, Julien
Choudhary, Himanshi
Cinque, Giorgio
Robert, Coralie
Bourdoncle, Anne
Picco, Raffaella
Mergny, Jean-Louis
Salgado, Gilmar F.
Xodo, Luigi E.
hnRNPA1/UP1 Unfolds KRAS G-Quadruplexes and Feeds a Regulatory Axis Controlling Gene Expression
title hnRNPA1/UP1 Unfolds KRAS G-Quadruplexes and Feeds a Regulatory Axis Controlling Gene Expression
title_full hnRNPA1/UP1 Unfolds KRAS G-Quadruplexes and Feeds a Regulatory Axis Controlling Gene Expression
title_fullStr hnRNPA1/UP1 Unfolds KRAS G-Quadruplexes and Feeds a Regulatory Axis Controlling Gene Expression
title_full_unstemmed hnRNPA1/UP1 Unfolds KRAS G-Quadruplexes and Feeds a Regulatory Axis Controlling Gene Expression
title_short hnRNPA1/UP1 Unfolds KRAS G-Quadruplexes and Feeds a Regulatory Axis Controlling Gene Expression
title_sort hnrnpa1/up1 unfolds kras g-quadruplexes and feeds a regulatory axis controlling gene expression
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8675163/
https://www.ncbi.nlm.nih.gov/pubmed/34926957
http://dx.doi.org/10.1021/acsomega.1c05538
work_keys_str_mv AT ferinoannalisa hnrnpa1up1unfoldskrasgquadruplexesandfeedsaregulatoryaxiscontrollinggeneexpression
AT marqueviellejulien hnrnpa1up1unfoldskrasgquadruplexesandfeedsaregulatoryaxiscontrollinggeneexpression
AT choudharyhimanshi hnrnpa1up1unfoldskrasgquadruplexesandfeedsaregulatoryaxiscontrollinggeneexpression
AT cinquegiorgio hnrnpa1up1unfoldskrasgquadruplexesandfeedsaregulatoryaxiscontrollinggeneexpression
AT robertcoralie hnrnpa1up1unfoldskrasgquadruplexesandfeedsaregulatoryaxiscontrollinggeneexpression
AT bourdoncleanne hnrnpa1up1unfoldskrasgquadruplexesandfeedsaregulatoryaxiscontrollinggeneexpression
AT piccoraffaella hnrnpa1up1unfoldskrasgquadruplexesandfeedsaregulatoryaxiscontrollinggeneexpression
AT mergnyjeanlouis hnrnpa1up1unfoldskrasgquadruplexesandfeedsaregulatoryaxiscontrollinggeneexpression
AT salgadogilmarf hnrnpa1up1unfoldskrasgquadruplexesandfeedsaregulatoryaxiscontrollinggeneexpression
AT xodoluigie hnrnpa1up1unfoldskrasgquadruplexesandfeedsaregulatoryaxiscontrollinggeneexpression