MMTV RNA packaging requires an extended long-range interaction for productive Gag binding to packaging signals.

IF 9.8 1区 生物学 Q1 Agricultural and Biological Sciences
PLoS Biology Pub Date : 2024-10-03 eCollection Date: 2024-10-01 DOI:10.1371/journal.pbio.3002827
Suresha G Prabhu, Vineeta N Pillai, Lizna Mohamed Ali, Valérie Vivet-Boudou, Akhil Chameettachal, Serena Bernacchi, Farah Mustafa, Roland Marquet, Tahir A Rizvi
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Abstract

The packaging of genomic RNA (gRNA) into retroviral particles relies on the specific recognition by the Gag precursor of packaging signals (Psi), which maintain a complex secondary structure through long-range interactions (LRIs). However, it remains unclear whether the binding of Gag to Psi alone is enough to promote RNA packaging and what role LRIs play in this process. Using mouse mammary tumor virus (MMTV), we investigated the effects of mutations in 4 proposed LRIs on gRNA structure and function. Our findings revealed the presence of an unsuspected extended LRI, and hSHAPE revealed that maintaining a wild-type-like Psi structure is crucial for efficient packaging. Surprisingly, filter-binding assays demonstrated that most mutants, regardless of their packaging capability, exhibited significant binding to Pr77Gag, suggesting that Gag binding to Psi is insufficient for efficient packaging. Footprinting experiments indicated that efficient RNA packaging is promoted when Pr77Gag binds to 2 specific sites within Psi, whereas binding elsewhere in Psi does not lead to efficient packaging. Taken together, our results suggest that the 3D structure of the Psi/Pr77Gag complex regulates the assembly of viral particles around gRNA, enabling effective discrimination against other viral and cellular RNAs that may also bind Gag efficiently.

MMTV RNA 包装需要延伸的长程相互作用,才能使 Gag 与包装信号有效结合。
将基因组 RNA(gRNA)包装成逆转录病毒颗粒依赖于 Gag 前体对包装信号(Psi)的特异性识别,包装信号通过长程相互作用(LRIs)维持复杂的二级结构。然而,Gag 与 Psi 的结合是否足以促进 RNA 包装,以及 LRIs 在这一过程中发挥什么作用,目前仍不清楚。我们利用小鼠乳腺肿瘤病毒(MMTV)研究了 4 个拟议的 LRIs 突变对 gRNA 结构和功能的影响。我们的发现揭示了一种未被发现的扩展 LRI 的存在,而 hSHAPE 则揭示了维持野生型 Psi 结构对于高效包装至关重要。令人惊讶的是,过滤结合实验表明,大多数突变体,无论其包装能力如何,都表现出与 Pr77Gag 的显著结合,这表明 Gag 与 Psi 的结合不足以实现高效包装。足迹实验表明,当 Pr77Gag 与 Psi 中的两个特定位点结合时,可促进有效的 RNA 包装,而与 Psi 中的其他位点结合则不会导致有效的包装。综上所述,我们的研究结果表明,Psi/Pr77Gag 复合物的三维结构调节了病毒颗粒围绕 gRNA 的组装,从而能够有效区分其他也可能与 Gag 有效结合的病毒和细胞 RNA。
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来源期刊
PLoS Biology
PLoS Biology BIOCHEMISTRY & MOLECULAR BIOLOGY-BIOLOGY
CiteScore
15.40
自引率
2.00%
发文量
359
审稿时长
3-8 weeks
期刊介绍: PLOS Biology is the flagship journal of the Public Library of Science (PLOS) and focuses on publishing groundbreaking and relevant research in all areas of biological science. The journal features works at various scales, ranging from molecules to ecosystems, and also encourages interdisciplinary studies. PLOS Biology publishes articles that demonstrate exceptional significance, originality, and relevance, with a high standard of scientific rigor in methodology, reporting, and conclusions. The journal aims to advance science and serve the research community by transforming research communication to align with the research process. It offers evolving article types and policies that empower authors to share the complete story behind their scientific findings with a diverse global audience of researchers, educators, policymakers, patient advocacy groups, and the general public. PLOS Biology, along with other PLOS journals, is widely indexed by major services such as Crossref, Dimensions, DOAJ, Google Scholar, PubMed, PubMed Central, Scopus, and Web of Science. Additionally, PLOS Biology is indexed by various other services including AGRICOLA, Biological Abstracts, BIOSYS Previews, CABI CAB Abstracts, CABI Global Health, CAPES, CAS, CNKI, Embase, Journal Guide, MEDLINE, and Zoological Record, ensuring that the research content is easily accessible and discoverable by a wide range of audiences.
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