Roles for mRNA methylation and lncRNA in Candida mating and pathogenicity

mRNA 甲基化和 lncRNA 在念珠菌交配和致病性中的作用

• 批准号: 9085341
• 负责人: Gary Guy Bushkin
• 金额: $ 5.8万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9085341
• 关键词: Adenosine; Animal Model; Antibodies; Antisense RNA; Benign; Biological Assay; Candida; Candida albicans; Cells; Cessation of life; Cis Tests; Consensus Sequence; Coupled; Cues; Data; Distant; Down-Regulation; Drug resistance; Eukaryota; Filament; Gene Expression Regulation; Genes; Genetic Variation; Goals; Growth; Health; High Pressure Liquid Chromatography; High-Throughput RNA Sequencing; Homologous Gene; Human; Immunocompromised Host; Individual; Infection; Life; Link; Mass Spectrum Analysis; Mating Types; Measures; Meiosis; Membrane; Messenger RNA; Methylation; Methyltransferase; Microbial Biofilms; Modification; Molecular; Molecular Genetics; Northern Blotting; Organism; Partner in relationship; Pathogenesis; Pathogenicity; Play; Process; Proteins; RNA; Regulation; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Saccharomyces cerevisiae; Skin; Starvation; Systemic infection; Testing; Transcript; Transplant Recipients; Untranslated RNA; Virulence; Yeasts; fungus; killings; methyl group; methylome; novel therapeutics; overexpression; pathogen; programs; promoter; response; transcriptome sequencing

DESCRIPTION (provided by applicant): This proposal is targeted at understanding the roles of mRNA methylation and long non-coding RNA, which are two molecular mechanisms of gene regulation, in Candida albicans virulence. C. albicans is a normally benign commensal, but can cause skin and mucosal membrane infections in healthy individuals, and life threatening systemic infections in susceptible individuals. C. albicans pathogenicity is linked to its ability o switch from growing in yeast-form cells to growing in filament and biofilm forms. While it cannot reproduce sexually, C. albicans can generate genetic diversity by mating, which has the potential to result in increased virulence and drug resistance. A better understanding of the molecular mechanisms behind these processes is required in order to identify new therapeutic avenues. mRNA methylation has been discovered decades ago, but the large scale of this modification in other eukaryotes has only recently became apparent, and its function in gene regulation is not yet understood. In Saccharomyces cerevisiae, IME4 encodes an mRNA methyltransferase, which plays a regulatory role in S. cerevisiae meiosis and filamentation. An IME4 homologue is present in C. albicans. Long non-coding RNA (lncRNA) are large transcripts that participate in gene regulation but do not encode any protein. S. cerevisiae IME4 expression is regulated by an antisense lncRNA, and this form of regulation appears to be conserved in C. albicans. Preliminary data indicates that mRNA methylation occurs in C. albicans, and that IME4 is expressed under filamentation inducing conditions. Furthermore, overexpression of IME4 inhibits filamentation. Using molecular genetics coupled with high throughput RNA sequencing, the proposed research will identify C. albicans IME4 mRNA targets, assess its role in mating, filamentation and biofilm formation, and study its regulation by lncRNA.

描述（申请人提供）：本提案旨在了解 mRNA 甲基化和长链非编码 RNA（基因调控的两种分子机制）在白色念珠菌毒力中的作用。 白色念珠菌通常是良性共生菌，但可引起健康个体的皮肤和粘膜感染，以及易感个体的危及生命的全身感染。白色念珠菌的致病性与其从酵母形式细胞中生长转变为丝状和生物膜形式生长的能力有关。虽然白色念珠菌不能有性繁殖，但它可以通过交配产生遗传多样性，这有可能导致毒力和耐药性增加。为了确定新的治疗途径，需要更好地了解这些过程背后的分子机制。 mRNA甲基化几十年前就被发现了，但这种修饰在其他真核生物中的大规模修饰直到最近才变得明显，其在基因调控中的功能尚不清楚。在酿酒酵母中，IME4 编码一种 mRNA 甲基转移酶，在酿酒酵母减数分裂和丝状化中发挥调节作用。 IME4 同源物存在于白色念珠菌中。长非编码RNA (lncRNA) 是参与基因调控但不编码任何蛋白质的大型转录物。酿酒酵母 IME4 表达受反义 lncRNA 调节，这种调节形式在白色念珠菌中似乎是保守的。 初步数据表明白色念珠菌中发生 mRNA 甲基化，并且 IME4 在丝状化诱导条件下表达。此外，IME4 的过度表达会抑制丝状形成。 拟议的研究将利用分子遗传学与高通量 RNA 测序相结合，识别白色念珠菌 IME4 mRNA 靶标，评估其在交配、丝状化和生物膜形成中的作用，并研究 lncRNA 对其的调节。