ERK3/MAPK6 dictates CDC42/RAC1 activity and ARP2/3-dependent actin polymerization

  1. Katarzyna Bogucka-Janczi
  2. Gregory Harms
  3. Marie-May Coissieux
  4. Mohamed Bentires-Alj
  5. Bernd Thiede
  6. Krishnaraj Rajalingam  Is a corresponding author
  1. Johannes Gutenberg University of Mainz, Germany
  2. University Hospital of Basel, Switzerland
  3. University of Basel, Switzerland
  4. University of Oslo, Norway

Abstract

The actin cytoskeleton is tightly controlled by RhoGTPases, actin binding-proteins and nucleation-promoting factors to perform fundamental cellular functions. We have previously shown that ERK3, an atypical MAPK controls IL-8 production and chemotaxis52. Here, we show in human cells that, ERK3 directly acts as a guanine nucleotide exchange factor for CDC42and phosphorylates the ARP3 subunit of the ARP2/3 complex at S418 to promote filopodia formation and actin polymerization, respectively. Consistently, depletion of ERK3 prevented both basal and EGF-dependent RAC1 and CDC42 activation, maintenance of F-actin content, filopodia formation and epithelial cell migration. Further, ERK3 protein bound directly to the purified ARP2/3 complex and augmented polymerization of actin in vitro. ERK3 kinase activity was required for the formation of actin-rich protrusions in mammalian cells. These findings unveil a fundamentally unique pathway employed by cells to control actin-dependent cellular functions.

Data availability

All source data files have been uploaded with the manuscript

Article and author information

Author details

  1. Katarzyna Bogucka-Janczi

    Cell Biology Unit, Johannes Gutenberg University of Mainz, Mainz, Germany
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6254-3359
  2. Gregory Harms

    Cell Biology Unit, Johannes Gutenberg University of Mainz, Mainz, Germany
    Competing interests
    No competing interests declared.
  3. Marie-May Coissieux

    University Hospital of Basel, Basel, Switzerland
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5017-5253
  4. Mohamed Bentires-Alj

    Department of Biomedicine, University of Basel, Basel, Switzerland
    Competing interests
    No competing interests declared.
  5. Bernd Thiede

    Department of Bioscience, University of Oslo, Oslo, Norway
    Competing interests
    No competing interests declared.
  6. Krishnaraj Rajalingam

    Cell Biology Unit, Johannes Gutenberg University of Mainz, Mainz, Germany
    For correspondence
    krishna@uni-mainz.de
    Competing interests
    Krishnaraj Rajalingam, KR is the founder and MD of KHR Biotec GmbH.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4175-9633

Funding

Else Kröner-Fresenius-Stiftung (SUN-MAPK)

  • Katarzyna Bogucka-Janczi
  • Gregory Harms

Deutsche Forschungsgemeinschaft (CRC1292)

  • Katarzyna Bogucka-Janczi
  • Krishnaraj Rajalingam

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: The animal experiments were performed as per the guideliens of University MEdical center Basel

Copyright

© 2023, Bogucka-Janczi et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 2,981
    views
  • 455
    downloads
  • 14
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Katarzyna Bogucka-Janczi
  2. Gregory Harms
  3. Marie-May Coissieux
  4. Mohamed Bentires-Alj
  5. Bernd Thiede
  6. Krishnaraj Rajalingam
(2023)
ERK3/MAPK6 dictates CDC42/RAC1 activity and ARP2/3-dependent actin polymerization
eLife 12:e85167.
https://doi.org/10.7554/eLife.85167

Share this article

https://doi.org/10.7554/eLife.85167

Further reading

    1. Cancer Biology
    Pierluigi Scerbo, Benjamin Tisserand ... Bertrand Ducos
    Research Article

    Why does a normal cell possibly harboring genetic mutations in oncogene or tumor suppressor genes becomes malignant and develops a tumor is a subject of intense debate. Various theories have been proposed but their experimental test has been hampered by the unpredictable and improbable malignant transformation of single cells. Here, using an optogenetic approach we permanently turn on an oncogene (KRASG12V) in a single cell of a zebrafish brain that, only in synergy with the transient co-activation of a reprogramming factor (VENTX/NANOG/OCT4), undergoes a deterministic malignant transition and robustly and reproducibly develops within 6 days into a full-blown tumor. The controlled way in which a single cell can thus be manipulated to give rise to cancer lends support to the ‘ground state theory of cancer initiation’ through ‘short-range dispersal’ of the first malignant cells preceding tumor growth.

    1. Cancer Biology
    Han V Han, Richard Efem ... Richard Z Lin
    Research Article

    Most human pancreatic ductal adenocarcinoma (PDAC) are not infiltrated with cytotoxic T cells and are highly resistant to immunotherapy. Over 90% of PDAC have oncogenic KRAS mutations, and phosphoinositide 3-kinases (PI3Ks) are direct effectors of KRAS. Our previous study demonstrated that ablation of Pik3ca in KPC (KrasG12D; Trp53R172H; Pdx1-Cre) pancreatic cancer cells induced host T cells to infiltrate and completely eliminate the tumors in a syngeneic orthotopic implantation mouse model. Now, we show that implantation of Pik3ca−/− KPC (named αKO) cancer cells induces clonal enrichment of cytotoxic T cells infiltrating the pancreatic tumors. To identify potential molecules that can regulate the activity of these anti-tumor T cells, we conducted an in vivo genome-wide gene-deletion screen using αKO cells implanted in the mouse pancreas. The result shows that deletion of propionyl-CoA carboxylase subunit B gene (Pccb) in αKO cells (named p-αKO) leads to immune evasion, tumor progression, and death of host mice. Surprisingly, p-αKO tumors are still infiltrated with clonally enriched CD8+ T cells but they are inactive against tumor cells. However, blockade of PD-L1/PD1 interaction reactivated these clonally enriched T cells infiltrating p-αKO tumors, leading to slower tumor progression and improve survival of host mice. These results indicate that Pccb can modulate the activity of cytotoxic T cells infiltrating some pancreatic cancers and this understanding may lead to improvement in immunotherapy for this difficult-to-treat cancer.