Deconstruction of the Ras switching cycle through saturation mutagenesis

Author:

Bandaru Pradeep123ORCID,Shah Neel H123ORCID,Bhattacharyya Moitrayee123,Barton John P4567ORCID,Kondo Yasushi123,Cofsky Joshua C123,Gee Christine L123ORCID,Chakraborty Arup K45678,Kortemme Tanja9,Ranganathan Rama101112,Kuriyan John12313ORCID

Affiliation:

1. Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, United States

2. California Institute for Quantitative Biosciences, University of California, Berkeley, Berkeley, United States

3. Howard Hughes Medical Institute, University of California, Berkeley, Berkeley, United States

4. Ragon Institute of MGH, MIT and Harvard, Cambridge, United States

5. Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, United States

6. Department of Physics, Massachusetts Institute of Technology, Cambridge, United States

7. Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, United States

8. Department of Chemistry, Massachusetts Institute of Technology, Cambridge, United States

9. Department of Bioengineering and Therapeutic Sciences, California Institute for Quantitative Biomedical Research, University of California, San Francisco, San Francisco, United States

10. Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, United States

11. Green Center for Systems Biology, University of Texas Southwestern Medical Center, Dallas, United States

12. Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, United States

13. Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, United States

Abstract

Ras proteins are highly conserved signaling molecules that exhibit regulated, nucleotide-dependent switching between active and inactive states. The high conservation of Ras requires mechanistic explanation, especially given the general mutational tolerance of proteins. Here, we use deep mutational scanning, biochemical analysis and molecular simulations to understand constraints on Ras sequence. Ras exhibits global sensitivity to mutation when regulated by a GTPase activating protein and a nucleotide exchange factor. Removing the regulators shifts the distribution of mutational effects to be largely neutral, and reveals hotspots of activating mutations in residues that restrain Ras dynamics and promote the inactive state. Evolutionary analysis, combined with structural and mutational data, argue that Ras has co-evolved with its regulators in the vertebrate lineage. Overall, our results show that sequence conservation in Ras depends strongly on the biochemical network in which it operates, providing a framework for understanding the origin of global selection pressures on proteins.

Funder

Howard Hughes Medical Institute

Damon Runyon Cancer Research Foundation

National Institutes of Health

Publisher

eLife Sciences Publications, Ltd

Subject

General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Medicine,General Neuroscience

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3