单分子光镊技术研究HIV-1逆转录酶的分子马达机制

逆转录是一种与多种疾病相关的重要生物现象，同时逆转录酶也是重要的分子生物学工具。虽然逆转录酶是重要的药物靶点，同时也广泛应用于分子生物学实验，但其作为分子马达的力学特性和运动机制一直未被阐明。本项目将利用光镊技术，研究单个HIV-1逆转录酶在不同核酸模板上的运动机制，测量其产生的力，并考察模板链上的折叠结构以及非模板互补链对逆转录过程的影响。本项目还将研究逆转录酶抑制剂对逆转录酶马达蛋白运动机制的调节作用。本项目研究成果将阐明HIV-1逆转录酶的力学特性，揭示其运动细节，从而为未来开发和优化抗逆转录药物，以及优化逆转录工具酶提供启示。

Reverse transcription is an important biological phenomenon relevant to many diseases. Reverse transcriptase is also an important molecular biological tool. Although reverse transcriptase is a critical drug target and widely used in molecular biology research, its mechanical and kinetic properties as a motor protein remain unclear. In this project, optical tweezers will be utilized to measure the motion and force of a single HIV-1 reverse transcriptase. Interactions between HIV-1 reverse transcriptase and mechanical obstacles, such as folded structures of the template strand and non-templated complementary strands, will be also investigated. Additionally, we will study effects of inhibitors on the motor activities of reverse transcriptase at the single-molecule level. Our studies will unravel the kinetic and mechanical details of the motor activities of HIV-1 reverse transcriptase, which will provide insights for antiretroviral drug development and for optimization of the tool enzymes for reverse transcription.