Frameshift mutations in the mmpR5 gene can have a bedaquiline-susceptible phenotype by retaining a protein structure and function similar to wild-type Mycobacterium tuberculosis.

IF 4.1 2区 医学 Q2 MICROBIOLOGY Antimicrobial Agents and Chemotherapy Pub Date : 2024-12-05 Epub Date: 2024-10-24 DOI:10.1128/aac.00854-24
J Snobre, C J Meehan, W Mulders, L Rigouts, R Buyl, B C de Jong, A Van Rie, O Tzfadia
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Abstract

Bedaquiline (BDQ) is crucial for the treatment of rifampicin-resistant tuberculosis, yet resistance threatens its effectiveness, mainly linked to mutations in the mmpR5 (Rv0678) gene. While frameshift mutations are thought to produce non-functional proteins, we hypothesize that they can result in conserved proteins through late-stop codons or alternative reading frames and remain BDQ susceptible. We extracted 512 isolates harboring frameshift mutations in mmpR5 from the World Health Organization (WHO) catalog and 68 isolates with minimum inhibitory concentration (MIC) in mycobacterial growth indicator tube (MGIT) through a literature review. Using BioPython and AlphaFold2 we computed open (ORF) and alternative reading frames (ARFs) sequences and protein structures and assessed similarity to the wild type using an alignment and template modeling (TM)-score. Among the WHO 512 isolates, 24.8% were BDQ-sensitive. Out of 184 unique frameshift mutations with available nucleotide information, a late-stop codon in the ORF occurred for 32% of the mutations. Also, 40.7% resulted in a conserved sequence, through the ORF or one of the forward ARFs. In 68 isolates with available MGIT MIC data, the presence of late-stop codons in the ORF (OR 4.71, 95% CI 1.36-19.3) or a conserved reading frame (OR 10.4, 95% CI 2.07-102.9) were associated with BDQ sensitivity. Protein structures from the conserved sequences showed high similarity (TM > 0.8). We show that frameshift mutations may retain BDQ susceptibility through late-stop codons in the ORF or conserved ARFs. These findings could improve the prediction of the BDQ phenotype from genomic data and have important implications for treatment decisions. Research Foundation-Flanders, Academy of Medical Sciences, the Wellcome Trust, the Government Department of Business, Energy and Industrial Strategy, the British Heart Foundation and Diabetes UK, and the Global Challenges Research Fund.IMPORTANCETuberculosis (TB), caused by Mycobacterium tuberculosis, remains the deadliest infectious disease and is particularly challenging to treat when it becomes drug-resistant. Bedaquiline (BDQ) is a recently recommended core drug for treating drug-resistant TB. However, resistance to bedaquiline is already emerging, primarily due to mutations in the mmpR5 gene. Identifying which mutations cause resistance and which do not is a critical knowledge gap. In particular, little is known about the effect of frameshift mutations, typically thought to make TB bacteria resistant to bedaquiline by producing non-functional proteins. Yet, one-quarter of isolates with a frameshift mutation are still susceptible to bedaquiline. How the bacteria produce a functional protein despite the frameshift mutation is unknown. We analyzed over 500 frameshift mutations using computational methods to model their effects on protein structure and bedaquiline resistance. Our findings revealed that some frameshift mutations can still produce functional proteins, allowing bacteria to remain sensitive to bedaquiline. Specifically, bacteria can produce a functional protein despite frameshift mutations if the mutation occurs near the end of the protein or if an alternative reading frame is available. These insights improve our ability to interpret mutations associated with bedaquiline, the most important drug for drug-resistant TB, allowing more accurate and effective treatment decisions.

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mmpR5 基因的框架移位突变可通过保留与野生型结核分枝杆菌相似的蛋白质结构和功能,产生贝达喹啉易感表型。
贝达喹啉(BDQ)是治疗耐利福平结核病的关键药物,但耐药性威胁着它的有效性,这主要与mmpR5(Rv0678)基因的突变有关。虽然换框突变被认为会产生无功能蛋白质,但我们假设它们可以通过晚期终止密码子或替代阅读框产生保守蛋白质,并保持对 BDQ 的敏感性。我们从世界卫生组织(WHO)的目录中提取了 512 个携带 mmpR5 框移突变的分离株,并通过文献回顾提取了 68 个在分枝杆菌生长指示管(MGIT)中具有最小抑菌浓度(MIC)的分离株。我们使用 BioPython 和 AlphaFold2 计算了开放(ORF)和替代阅读框(ARF)序列和蛋白质结构,并使用比对和模板建模(TM)分数评估了与野生型的相似性。在世界卫生组织的 512 个分离株中,24.8% 对 BDQ 敏感。在184个有核苷酸信息的独特换框突变中,32%的突变发生在ORF中的晚停密码子上。此外,40.7%的突变是通过 ORF 或一个前向 ARF 的保守序列导致的。在有 MGIT MIC 数据的 68 个分离株中,ORF 中存在晚终止密码子(OR 4.71,95% CI 1.36-19.3)或保守阅读框(OR 10.4,95% CI 2.07-102.9)与 BDQ 敏感性相关。来自保守序列的蛋白质结构显示出高度相似性(TM > 0.8)。我们的研究表明,换框突变可能通过ORF或保守的ARF中的晚期终止密码子保持对BDQ的敏感性。这些发现可以改善从基因组数据中对 BDQ 表型的预测,并对治疗决策产生重要影响。该研究由弗兰德斯研究基金会、医学科学院、惠康基金会、英国商业、能源和工业战略部、英国心脏基金会、英国糖尿病基金会以及全球挑战研究基金资助。贝达喹啉(BDQ)是最近推荐的治疗耐药性结核病的核心药物。然而,贝达喹啉的耐药性已经出现,主要是由于 mmpR5 基因突变造成的。确定哪些突变会导致耐药性,哪些不会,是一个关键的知识缺口。尤其是对换框突变的影响知之甚少,换框突变通常被认为是通过产生无功能蛋白质使结核菌对贝达喹啉产生耐药性。然而,四分之一发生了框架转换突变的分离株仍然对贝达喹啉敏感。这些细菌是如何在发生移帧突变的情况下产生功能性蛋白质的,目前尚不清楚。我们使用计算方法分析了 500 多种框架转换突变,以模拟它们对蛋白质结构和贝达喹啉抗性的影响。我们的研究结果表明,一些换框突变仍能产生功能蛋白,使细菌对贝达喹啉保持敏感。具体来说,如果突变发生在蛋白质末端附近或存在替代阅读框,那么尽管存在换框突变,细菌仍能产生功能性蛋白质。这些见解提高了我们解释与贝达喹啉(治疗耐药性结核病的最重要药物)相关的突变的能力,从而可以做出更准确、更有效的治疗决定。
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来源期刊
CiteScore
10.00
自引率
8.20%
发文量
762
审稿时长
3 months
期刊介绍: Antimicrobial Agents and Chemotherapy (AAC) features interdisciplinary studies that build our understanding of the underlying mechanisms and therapeutic applications of antimicrobial and antiparasitic agents and chemotherapy.
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