bHLH Proteins and Neural Development in C. elegans

真核生物的发育依赖于三个精心安排的过程：分化、形态发生和生长。为了做到这一点，发育中的生物体负责调节这些事件所需的基因的空间和时间表达。基本的螺旋环抑制（bHLH）转录因子家族调节许多这些基因的表达，通常通过依赖于对环境提供的感觉信号的适当反应的调节级联。约翰逊实验室的成员对了解bHLH蛋白家族在线虫（秀丽隐杆线虫）神经发育中的特定作用感兴趣。在之前NSF支持的研究中，研究生和本科生确定并启动了一个bHLH基因hlh-17的分子研究。本研究旨在通过以下两个具体目标更清楚地描述hlh-17的表达和生物活性：（1）使用下拉测定和酵母双杂交系统鉴定能够与HLH-17相互作用和/或影响HLH-17的其它bHLH蛋白，和（2）确定hLH-17是否影响，或受其影响，通过定量RT-PCR检测调节衰老和寿命的基因。 本研究为进一步了解HLH-17在C. elegans，提供信息的哺乳动物同源物的HLH-17在哺乳动物神经发育的可能作用，并提供进一步了解少数bHLH蛋白，被认为是作为一个负的，非DNA结合转录调节。该项目的更广泛的影响是，所有的研究将在摩根州立大学，一个历史上的黑人机构位于马里兰州的巴尔的摩市，由本科生研究人员和硕士学位候选人（其中大多数将是代表性不足的少数民族或妇女），所有这些人都将参加当地，国家和国际研究研讨会。 此外，约翰逊实验室有一个长期的传统，即让所有级别（K-12）的学生参与研究活动，通过本科生在当地小学进行的演示，通过为期4-6周的PI赞助的中学生实验室实验，以及通过高中/中学生实习，这一传统将在本项目中继续下去。

Eukaryotic development depends on three carefully orchestrated processes: differentiation, morphogenesis, and growth. In order to do so, the developing organism is responsible for regulating the spatial and temporal expression of the genes that are required for these events. The basic Helix-Loop-Helix (bHLH) transcription factor family regulates the expression of a number of these genes, often through regulatory cascades that rely on the proper responses to sensory signals provided by the environment. Members of the Johnson laboratory are interested in understanding the specific role of the bHLH family of proteins on neural development in the roundworm, Caenorhabditis elegans. In a previous NSF-supported study, graduate and undergraduate students identified and initiated a molecular study of one bHLH gene, hlh-17. The research to take place in the present study is designed to delineate more clearly the expression and biological activity of hlh-17 through two specific aims: (1) identify other bHLH proteins that are capable of interacting with and/or influencing HLH-17 using pull-down assays and the yeast two-hybrid system, and (2) determine if hlh-17 influences, or is influenced by, genes that regulate aging and longevity by using quantitative RT-PCR. This project should provide further insight into the role of HLH-17 in neurological behaviors of C. elegans, provide information on the possible roles of the mammalian homologs of HLH-17 in mammalian neural development, and provide further insight into one of the few bHLH proteins that is believed to act as a negative, non-DNA binding regulator of transcription. The broader impact of the project is that all of the research will be done at Morgan State University, a historically black institution located in Baltimore City, Maryland, by undergraduate student researchers and a Master's degree candidate (most of whom will be underrepresented minorities or women), all of whom will participate in local, national, and international research symposia. In addition, the Johnson laboratory has a long-standing tradition of involving students at all levels (K-12) in the research activities, through undergraduate student-led demonstrations at local elementary schools, through 4-6 week long PI-sponsored laboratory experiments with middle school students, and through high school/middle school student internships, a tradition that will continue with this project.