Mutational Pleiotropy, Epistasis, and the Adaptive Evolution of Hemoglobin Function

突变多效性、上位性和血红蛋白功能的适应性进化

• 批准号: 10246848
• 负责人: Jay Storz
• 金额: $ 36.97万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-22 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10246848
• 关键词: 3-Dimensional; Address; Allosteric Regulation; Amino Acids; Bicarbonates; Biochemical; Biology; Biophysical Process; Biophysics; Cells; Crystallization; Diving; Erythrocytes; Evolution; Genetic; Genetic Epistasis; Goals; Hemoglobin; Individual; Ions; Light; Metabolic; Minor; Molecular; Molecular Evolution; Mutation; Nature; Outcome; Pathway interactions; Phenotype; Physiological; Physiology; Prevalence; Property; Protein Engineering; Proteins; Recombinants; Recording of previous events; Regulation; Research Project Grants; Resolution; Role; Site; Site-Directed Mutagenesis; Structure; System; Vertebrates; X-Ray Crystallography; base; causal variant; combinatorial; design; experimental study; innovation; inorganic phosphate; insight; mutant; novel; oxygen transport; pleiotropism; protein function; sample fixation; structural biology; transfusion medicine

The step-by-step evolution of novel phenotypes is central to several fundamental questions in biology. In studies of novel protein functions, the problem becomes experimentally tractable if it is possible to identify and functionally characterize the complete set of causative mutations. With such a system, it is possible to address key questions: Do novel functions evolve via the successive fixation of beneficial mutations that each produce an adaptive change in phenotype when they first arise? Alternatively, are evolutionary transitions in protein function facilitated by neutral mutations that produce no adaptive benefit when they first arise, but which potentiate the function-altering effects of subsequent mutations? By reconstructing all possible mutational pathways that connect ancestral and descendant proteins it is also possible to address fundamental questions about the roles of contingency and determinism in protein evolution. For example: Can novel functions evolve from any possible ancestral starting point, or are specific evolutionary outcomes contingent on prior history? We will address these questions by experimentally dissecting the molecular basis of a key physiological innovation during vertebrate evolution. Specifically, we will examine the evolution of a unique allosteric mechanism for regulating hemoglobin (Hb) function in the red blood cells of crocodilians. This unique mode of allosteric regulatory control contributes to crocodilians’ extraordinary capacities for breath-hold diving. Using ancestral protein resurrection in conjunction with a combinatorial protein engineering approach based on site- directed mutagenesis, we will examine the effects of sequential mutational steps in the evolution of the novel allosteric mechanism of crocodilian Hb. We will also obtain insights into the structural basis of the change in Hb function, as X-ray crystallography experiments will reveal biophysical mechanisms at atomic resolution. The specific aims of the project are as follows: (1) Identify the specific mutations that are responsible for the evolution of the novel protein function, and quantify their additive and nonadditive effects; and (2) Identify and characterize the biophysical mechanisms responsible for the functional transition (gain of novel function, loss of ancestral function). In combination, accomplishing Specific Aims 1 and 2 will reveal the molecular basis of a key physiological innovation and will provide general insights into the pathways by which such innovations evolve.

新表型的逐步进化是生物学中几个基本问​​题的核心。 研究新的蛋白质功能，如果可以识别和解决这个问题，那么这个问题就可以通过实验解决。 通过这样的系统，可以在功能上表征完整的致病突变。 关键问题：新功能是否通过连续固定每个产生的有益突变而进化？ 或者，是蛋白质的进化转变吗？ 中性突变促进的功能，中性突变首次出现时不会产生适应性益处，但 通过重建所有可能的突变来增强后续突变的功能改变效应？ 连接祖先和后代蛋白质的途径也有可能解决基本问题 关于偶然性和决定论在蛋白质进化中的作用，例如：新功能能否进化。 从任何可能的祖先起点开始，还是特定的进化结果取决于先前的历史？ 我们将通过实验剖析关键生理学的分子基础来解决这些问题 具体来说，我们将研究独特的变构的进化。 调节鳄鱼红细胞中血红蛋白（Hb）功能的机制是这种独特的模式。 变构调节控制有助于鳄鱼具有非凡的屏气潜水能力。 祖先蛋白质复活与基于位点的组合蛋白质工程方法相结合 定向诱变，我们将研究连续突变步骤对小说进化的影响 我们还将深入了解 Hb 变化的结构基础。 功能，因为 X 射线晶体学实验将揭示原子分辨率的生物物理机制。 该项目的具体目标如下：（1）确定导致该现象的具体突变。 新蛋白质功能的进化，并量化其加性和非加性效应；以及 (2) 识别和 描述负责功能转变的生物物理机制（新功能的获得、丧失 结合起来，实现具体目标 1 和 2 将揭示 a 的分子基础。 关键的生理学创新，并将提供对此类创新途径的一般见解 发展。

项目成果

Experimental approaches to evaluate the contributions of candidate protein-coding mutations to phenotypic evolution.

评估候选蛋白质编码突变对表型进化的贡献的实验方法。

• DOI: 10.1007/978-1-61779-228-1_22
• 发表时间: 2011
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Storz,JayF;Zera,AnthonyJ
• 通讯作者: Zera,AnthonyJ

Epistasis constrains mutational pathways of hemoglobin adaptation in high-altitude pikas.

• DOI: 10.1093/molbev/msu311
• 发表时间: 2015-02
• 期刊: Molecular biology and evolution
• 影响因子: 10.7
• 作者: Tufts DM;Natarajan C;Revsbech IG;Projecto-Garcia J;Hoffmann FG;Weber RE;Fago A;Moriyama H;Storz JF
• 通讯作者: Storz JF

Oxygenation properties and oxidation rates of mouse hemoglobins that differ in reactive cysteine content.

• DOI: 10.1016/j.cbpa.2011.11.004
• 发表时间: 2012-02
• 期刊: COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY A-MOLECULAR & INTEGRATIVE PHYSIOLOGY
• 影响因子: 2.3
• 作者: Storz, Jay F.;Weber, Roy E.;Fago, Angela
• 通讯作者: Fago, Angela

Evolution of physiological performance capacities and environmental adaptation: insights from high-elevation deer mice (Peromyscus maniculatus)

• DOI: 10.1093/jmammal/gyy173
• 发表时间: 2019-05-23
• 期刊: JOURNAL OF MAMMALOGY
• 影响因子: 1.7
• 作者: Storz, Jay F.;Cheviron, Zachary A.;Scott, Graham R.
• 通讯作者: Scott, Graham R.

Intraspecific polymorphism, interspecific divergence, and the origins of function-altering mutations in deer mouse hemoglobin.

• DOI: 10.1093/molbev/msu403
• 发表时间: 2015-04
• 期刊: Molecular biology and evolution
• 影响因子: 10.7
• 作者: Natarajan C;Hoffmann FG;Lanier HC;Wolf CJ;Cheviron ZA;Spangler ML;Weber RE;Fago A;Storz JF
• 通讯作者: Storz JF