Cross-species analysis of LZTR1 loss-of-function mutants demonstrates dependency to RIT1 orthologs

  1. Antonio Cuevas-Navarro
  2. Laura Rodriguez-Muñoz
  3. Joaquim Grego-Bessa
  4. Alice Cheng
  5. Katherine A Rauen
  6. Anatoly Urisman
  7. Frank McCormick
  8. Gerardo Jimenez
  9. Pau Castel  Is a corresponding author
  1. University of California, San Francisco, United States
  2. Consejo Superior de Investigaciones Científicas, Spain
  3. Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), Spain
  4. University of California, Davis, United States
  5. New York University, United States

Abstract

RAS GTPases are highly conserved proteins involved in the regulation of mitogenic signaling. We have previously described a novel Cullin 3 RING E3 ubiquitin ligase complex formed by the substrate adaptor protein LZTR1 that binds, ubiquitinates, and promotes proteasomal degradation of the RAS GTPase RIT1. In addition, others have described that this complex is also responsible for the ubiquitination of classical RAS GTPases. Here, we have analyzed the phenotypes of Lztr1 loss-of-function mutants in both fruit flies and mice and have demonstrated a biochemical preference for their RIT1 orthologs. Moreover, we show that Lztr1 is haplosufficient in mice and that embryonic lethality of the homozygous null allele can be rescued by deletion of Rit1. Overall, our results indicate that, in model organisms, RIT1 orthologs are the preferred substrates of LZTR1.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting file; Source Data files have been provided for all Figures.

Article and author information

Author details

  1. Antonio Cuevas-Navarro

    Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
  2. Laura Rodriguez-Muñoz

    Institute for Molecular Biology of Barcelona, Consejo Superior de Investigaciones Científicas, Barcelona, Spain
    Competing interests
    No competing interests declared.
  3. Joaquim Grego-Bessa

    Intercellular Signalling in Cardiovascular Development and Disease Laboratory, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), Madrid, Spain
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0938-2346
  4. Alice Cheng

    Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
  5. Katherine A Rauen

    MIND Institute, University of California, Davis, Sacramento, United States
    Competing interests
    No competing interests declared.
  6. Anatoly Urisman

    Department of Anatomic Pathology, University of California, San Francisco, San Francisco, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8364-5303
  7. Frank McCormick

    Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, United States
    Competing interests
    Frank McCormick, is a consultant for Ideaya Biosciences, Kura Oncology, Leidos Biomedical Research, Pfizer, Daiichi Sankyo, Amgen, PMV Pharma, OPNA-IO, and Quanta Therapeutics and has received research grants from Boehringer-Ingelheim and is a consultant for and cofounder of BridgeBio Pharma..
  8. Gerardo Jimenez

    Institute for Molecular Biology of Barcelona, Consejo Superior de Investigaciones Científicas, Barcelona, Spain
    Competing interests
    No competing interests declared.
  9. Pau Castel

    Department of Biochemistry and Molecular Pharmacology, New York University, New York, United States
    For correspondence
    pau.castel@nyulangone.org
    Competing interests
    Pau Castel, PC is a founder and advisory board of Venthera..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4972-4347

Funding

National Cancer Institute (F31CA265066)

  • Antonio Cuevas-Navarro

National Cancer Institute (R35CA197709)

  • Frank McCormick

National Cancer Institute (R00CA245122)

  • Pau Castel

DOD CDMRP Neurofibromatosis Research Program (W81XWH-20-1-0391)

  • Pau Castel

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

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols (#AN165444 and #AN179937) of the University of California San Francisco.

Copyright

© 2022, Cuevas-Navarro 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

  • 1,741
    views
  • 270
    downloads
  • 12
    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. Antonio Cuevas-Navarro
  2. Laura Rodriguez-Muñoz
  3. Joaquim Grego-Bessa
  4. Alice Cheng
  5. Katherine A Rauen
  6. Anatoly Urisman
  7. Frank McCormick
  8. Gerardo Jimenez
  9. Pau Castel
(2022)
Cross-species analysis of LZTR1 loss-of-function mutants demonstrates dependency to RIT1 orthologs
eLife 11:e76495.
https://doi.org/10.7554/eLife.76495

Share this article

https://doi.org/10.7554/eLife.76495

Further reading

    1. Cancer Biology
    Yumin Fu, Xinyu Guo ... Lianxin Liu
    Review Article

    Hepatocellular carcinoma (HCC), the most common type of liver tumor, is a leading cause of cancer-related deaths, and the incidence of liver cancer is still increasing worldwide. Curative hepatectomy or liver transplantation is only indicated for a small population of patients with early-stage HCC. However, most patients with HCC are not candidates for radical resection due to disease progression, leading to the choice of the conventional tyrosine kinase inhibitor drug sorafenib as first-line treatment. In the past few years, immunotherapy, mainly immune checkpoint inhibitors (ICIs), has revolutionized the clinical strategy for HCC. Combination therapy with ICIs has proven more effective than sorafenib, and clinical trials have been conducted to apply these therapies to patients. Despite significant progress in immunotherapy, the molecular mechanisms behind it remain unclear, and immune resistance is often challenging to overcome. Several studies have pointed out that the complex intercellular communication network in the immune microenvironment of HCC regulates tumor escape and drug resistance to immune response. This underscores the urgent need to analyze the immune microenvironment of HCC. This review describes the immunosuppressive cell populations in the immune microenvironment of HCC, as well as the related clinical trials, aiming to provide insights for the next generation of precision immunotherapy.

    1. Cancer Biology
    2. Genetics and Genomics
    Li Min, Fanqin Bu ... Shutian Zhang
    Research Article

    It takes more than 20 years for normal colorectal mucosa to develop into metastatic carcinoma. The long time window provides a golden opportunity for early detection to terminate the malignant progression. Here, we aim to enable liquid biopsy of T1a stage colorectal cancer (CRC) and precancerous advanced adenoma (AA) by profiling circulating small extracellular vesicle (sEV)-derived RNAs. We exhibited a full RNA landscape for the circulating sEVs isolated from 60 participants. A total of 58,333 annotated RNAs were detected from plasma sEVs, among which 1,615 and 888 sEV-RNAs were found differentially expressed in plasma from T1a stage CRC and AA compared to normal controls (NC). Then we further categorized these sEV-RNAs into six modules by a weighted gene coexpression network analysis and constructed a 60-gene t-SNE model consisting of the top 10 RNAs of each module that could well distinguish T1a stage CRC/AA from NC samples. Some sEV-RNAs were also identified as indicators of specific endoscopic and morphological features of different colorectal lesions. The top-ranked biomarkers were further verified by RT-qPCR, proving that these candidate sEV-RNAs successfully identified T1a stage CRC/AA from NC in another cohort of 124 participants. Finally, we adopted different algorithms to improve the performance of RT-qPCR-based models and successfully constructed an optimized classifier with 79.3% specificity and 99.0% sensitivity. In conclusion, circulating sEVs of T1a stage CRC and AA patients have distinct RNA profiles, which successfully enable the detection of both T1a stage CRC and AA via liquid biopsy.