NUCLEOTIDE SYNTHESIS AND METABOLISM IN MYCOBACTERIA

分枝杆菌中的核苷酸合成和代谢

• 批准号: 6256350
• 负责人: Ananda Mohan Chakrabarty
• 金额: $ 19.41万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2003-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6256350
• 关键词: Mycobacterium bovis; bacterial genetics; enzyme activity; gene deletion mutation; microorganism culture; microorganism metabolism; molecular cloning; nucleic acid biosynthesis; nucleic acid sequence; nucleoside diphosphate kinase; nucleotide metabolism; nucleotides; ovalbumin; receptor; receptor binding; site directed mutagenesis

DESCRIPTION (Adapted from the Applicant's Abstract): Many of the mycobacterial cell wall polymers are derived from basic building blocks of nucleoside diphosphate-sugar residues such as GDP-mannose and UDP-galactose which require nucleoside triphosphates (NTPs) such as GTP or UTP for each molecule of the sugar residue present in the polymer. Nothing is known about how the precursor NTP pool in the mycobacterial cells is regulated. The Investigator has recently isolated 4 different proteins from Mycobacterium smegmatis that modulate the specificity of nucleoside diphosphate kinase (Ndk) to primarily synthesize either GTP, or UTP, or CTP. Whether such proteins may play a role in supplying specific NTPs for cell wall/envelope components is not know at present. The investigators propose to characterize Ndk-complexing proteins in M. bovis-BCG, isolate their genes, and attempt to make knockout mutations to examine if loss of the gene functions may result in altered cell wall synthesis as characterized chemically. They have also recently reported the secretability of two ATP-utilizing enzymes (Ndk and ATPase) of M. bovis BCG. The secretion of these two enzymes is facilitated in the presence of eukaryotic proteins such as bovine serum albumin or ovalbumin. Preliminary evidence suggests that the presence of receptors on the surface of M. bovis BCG cells that bind ovalbumin. They propose to isolate and characterize these receptors. Additionally, they would like to make deletion and site-directed mutations in the ndk gene of M. bovis BCG to examine the role of putative sequences (motifs) in the secretability of Ndk.

描述（改编自申请人的摘要）：许多分枝杆菌属的细菌， 细胞壁聚合物衍生自核苷的基本结构单元 二磷酸-糖残基，如GDP-甘露糖和UDP-半乳糖，其需要 核苷三磷酸（NTP），如GTP或UTP，用于每个分子的药物组合物。 聚合物中存在的糖残基。我们不知道前体是如何 分枝杆菌细胞中的NTP库受到调节。调查人员最近 从耻垢分枝杆菌中分离出4种不同的蛋白质， 核苷二磷酸激酶（Ndk）对主要合成 GTP、UTP或CTP。这些蛋白质是否可以在供应 目前还不知道细胞壁/被膜组分的具体NTPs。的 研究者们提出在M.牛-BCG， 分离他们的基因，并试图使敲除突变，以检查是否损失 基因功能的改变可能导致细胞壁合成的改变， 以化学方式表征。他们最近还报道了 M.牛卡介苗的分泌 这两种酶在真核生物蛋白质的存在下被促进， 牛血清白蛋白或卵清蛋白。初步证据表明， M.结合卵白蛋白的牛卡介苗细胞。 他们建议分离和表征这些受体。而且两个人 希望对M. 牛BCG检查推定序列（基序）在 Ndk的分泌性