CAREER: Understanding the Molecular Mechanisms of Insect Cuticular Chitin Maintenance

职业:了解昆虫表皮几丁质维持的分子机制

基本信息

  • 批准号:
    2338209
  • 负责人:
  • 金额:
    $ 123万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
    Continuing Grant
  • 财政年份:
    2024
  • 资助国家:
    美国
  • 起止时间:
    2024-03-01 至 2029-02-28
  • 项目状态:
    未结题

项目摘要

Insects and other arthropods have a hard exoskeleton or cuticle, which must be shed in a process called molting for the animal to grow. This NSF CAREER Award aims to investigate mechanisms involved in insect cuticle formation by testing three possible mechanisms whereby the newly synthesized cuticle is protected from degradation during molting as the old cuticle is broken down by enzymes. While insects provide numerous benefits to humankind, they can also inflict extensive damage to crops and serve as vectors for pathogens that cause deadly diseases. Therefore, the research results could lead to innovative pest and vector management strategies. The acquired knowledge will be integrated into existing curricula and active learning approaches involving 3-D visualization modules on insect physiology will be developed, thereby enhancing the educational impact of the project. The project will offer multidisciplinary training to undergraduate and graduate students in the fields of insect physiology, biochemistry, molecular biology, and bioinformatics. By providing underprivileged and underrepresented minority female undergraduate students from local institutions with comprehensive laboratory research experiences and opportunities to present their work at regional/national conferences, the project seeks to empower those students to pursue STEM-based careers.Chitin serves as a crucial structural component of the insect cuticle (exoskeleton), peritrophic matrix (gut lining), trachea, and eggshell. Insects periodically replace their old cuticle through molting or ecdysis to accommodate growth. During each molt, a new cuticle forms, while the old one simultaneously undergoes partial degradation by chitinases before being shed. Notably, chitinases do not degrade the new cuticular chitin despite being abundant in the procuticle. Previous research has demonstrated the critical role of the protein Knickkopf (Knk) in preventing premature degradation of the new cuticle by chitinases, albeit through an unknown mechanism. This CAREER project investigates how newly synthesized cuticular chitin is shielded from chitinase activity during molting to ensure the integrity of the newly forming cuticle. It will test the hypothesis that chitin deacetylases play a pivotal role in maintaining cuticle integrity by shielding against chitinases, potentially in collaboration with Knk. The project will (1) examine the impact of chitin deacetylation on chitinase function, (2) identify accessory proteins required for chitin deacetylase function, and (3) assess the importance of chitin deacetylation in gut peritrophic matrix integrity. To better understand the function of chitin deacetylases in maintaining cuticle integrity, the research will employ, and train students in, various molecular biology, confocal microscopy, transmission electron microscopy and mass-spectrometry techniques.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
昆虫和其他节肢动物具有坚硬的外骨骼或角质层,必须在称为蜕皮的过程中脱落才能使动物生长。该 NSF 职业奖旨在通过测试三种可能的机制来研究昆虫角质层形成的机制,即旧角质层被酶分解时,新合成的角质层在蜕皮过程中免受降解。虽然昆虫为人类带来了许多好处,但它们也可能对农作物造成广泛损害,并成为导致致命疾病的病原体的载体。因此,研究结果可能会带来创新的害虫和病媒管理策略。所获得的知识将整合到现有课程中,并将开发涉及昆虫生理学 3D 可视化模块的主动学习方法,从而增强该项目的教育影响。该项目将为昆虫生理学、生物化学、分子生物学和生物信息学领域的本科生和研究生提供多学科培训。通过为当地机构的贫困和代表性不足的少数族裔女本科生提供全面的实验室研究经验和在区域/国家会议上展示其工作的机会,该项目旨在使这些学生能够追求基于 STEM 的职业。甲壳素是一个重要的结构组成部分昆虫角质层(外骨骼)、围食基质(肠壁)、气管和蛋壳。昆虫通过蜕皮或蜕皮定期更换旧角质层以适应生长。在每次蜕皮过程中,都会形成新的角质层,而旧的角质层在脱落之前同时会被几丁质酶部分降解。值得注意的是,尽管前角质中含量丰富,几丁质酶不会降解新的角质层几丁质。先前的研究已经证明了蛋白质 Knickkopf (Knk) 在防止几丁质酶过早降解新角质层方面发挥着关键作用,尽管其机制未知。该职业项目研究了新合成的角质层几丁质在蜕皮过程中如何免受几丁质酶活性的影响,以确保新形成的角质层的完整性。它将检验甲壳质脱乙酰酶通过屏蔽几丁质酶在维持角质层完整性方面发挥关键作用的假设,可能与 Knk 合作。该项目将(1)检查几丁质脱乙酰化对几丁质酶功能的影响,(2)鉴定几丁质脱乙酰化酶功能所需的辅助蛋白,以及(3)评估几丁质脱乙酰化在肠道围营养基质完整性中的重要性。为了更好地了解几丁质脱乙酰酶在维持角质层完整性方面的功能,该研究将使用各种分子生物学、共聚焦显微镜、透射电子显微镜和质谱技术,并对学生进行培训。该奖项反映了 NSF 的法定使命,并被认为是值得的通过使用基金会的智力优势和更广泛的影响审查标准进行评估来获得支持。

项目成果

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