Modified nucleotide cofactors and the interaction of the RNAome with the proteome

修饰的核苷酸辅因子以及 RNAome 与蛋白质组的相互作用

• 批准号: 8337709
• 负责人: GARY B RUVKUN
• 金额: $ 34.82万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-30 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8337709
• 关键词: Affect; Amino Acids; Animals; Bar Codes; Base Pairing; Binding; Biology; Caenorhabditis elegans; Charge; Chemicals; Cloning; Code; Coenzyme A; Comparative Genomic Analysis; Complement; Complementary RNA; Coupled; DNA; Data; Defect; Disulfides; Eukaryotic Cell; Fostering; Genome; Histone H3; Libraries; Ligation; Link; Maps; Mass Spectrum Analysis; Mediating; Methionine; MicroRNAs; Modeling; Modification; Nucleic Acid Binding; Nucleic Acid Hybridization; Nucleic Acids; Nucleotides; Oligonucleotides; Pathway interactions; Peptide Fragments; Peptide Hydrolases; Peptides; Plants; Positioning Attribute; Protein Analysis; Protein Biochemistry; Proteins; Proteome; Proteomics; RNA; RNA Binding; RNA Interference; Research; Series; Small Interfering RNA; Small RNA; Surveys; Techniques; Temperature; Tertiary Protein Structure; Tetrahymena; alpha Tubulin; cofactor; comparative; inorganic phosphate; link protein; melting; nuclease; sulfation

DESCRIPTION (provided by applicant): We have preliminary evidence that small RNAs of the size of siRNA and miRNAs are coupled to modified nucleotides such as coenzyme A and bis-MGD. These modifications would allow small RNAs to covalently engage with the proteome to constitute nucleic acid bar codes on these proteins. We propose to use powerful nucleic acid hybridization and selection techniques to purify the proteins covalently linked to RNA and to use mass spectroscopy to identify those linked proteins. Using RNAi of these proteins we will survey for defects in small RNA function and thus an involvement of these proteins in the functions of small RNAs. Because all of the modifications are predicted to occur at the 5' end of small RNAs, most of these modified RNAs will have been systematically missed by surveys most of which demands 5' OH or 5' phosphates on the RNAs for cloning.

描述（由申请人提供）：我们有初步证据表明，siRNA和miRNA大小的小RNA与辅酶A和BIS-MGD等修饰的核苷酸耦合。这些修饰将使小的RNA与蛋白质组共价互动，以构成这些蛋白质上的核酸条代码。我们建议使用强大的核酸杂交和选择技术来纯化与RNA共价关联的蛋白质，并使用质谱识别那些链接的蛋白质。使用这些蛋白质的RNAI，我们将调查小RNA功能中的缺陷，从而参与这些蛋白质中的小RNA功能。由于预计所有修改都会发生在小RNA的5'末端，因此大多数修改的RNA将被调查系统地遗漏，其中大多数要求在RNA上进行5'OH或5'磷酸盐以进行克隆。