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1. Limits to detecting epistasis in the fitness landscape of HIV

   https://doi.org/10.1371/journal.pone.0262314  

   Biswas, Avik; Haldane, Allan; Levy, Ronald M. (January 2022, PLOS ONE)

   Gallicchio, Emilio (Ed.)

   The rapid evolution of HIV is constrained by interactions between mutations which affect viral fitness. In this work, we explore the role of epistasis in determining the mutational fitness landscape of HIV for multiple drug target proteins, including Protease, Reverse Transcriptase, and Integrase. Epistatic interactions between residues modulate the mutation patterns involved in drug resistance, with unambiguous signatures of epistasis best seen in the comparison of the Potts model predicted and experimental HIV sequence “prevalences” expressed as higher-order marginals (beyond triplets) of the sequence probability distribution. In contrast, experimental measures of fitness such as viral replicative capacities generally probe fitness effects of point mutations in a single background, providing weak evidence for epistasis in viral systems. The detectable effects of epistasis are obscured by higher evolutionary conservation at sites. While double mutant cycles in principle, provide one of the best ways to probe epistatic interactions experimentally without reference to a particular background, we show that the analysis is complicated by the small dynamic range of measurements. Overall, we show that global pairwise interaction Potts models are necessary for predicting the mutational landscape of viral proteins. 

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2. Mutational robustness changes during long-term adaptation in laboratory budding yeast populations

   https://doi.org/10.7554/eLife.76491  

   Johnson, Milo S; Desai, Michael M (July 2022, eLife)

   As an adapting population traverses the fitness landscape, its local neighborhood (i.e., the collection of fitness effects of single-step mutations) can change shape because of interactions with mutations acquired during evolution. These changes to the distribution of fitness effects can affect both the rate of adaptation and the accumulation of deleterious mutations. However, while numerous models of fitness landscapes have been proposed in the literature, empirical data on how this distribution changes during evolution remains limited. In this study, we directly measure how the fitness landscape neighborhood changes during laboratory adaptation. Using a barcode-based mutagenesis system, we measure the fitness effects of 91 specific gene disruption mutations in genetic backgrounds spanning 8000–10,000 generations of evolution in two constant environments. We find that the mean of the distribution of fitness effects decreases in one environment, indicating a reduction in mutational robustness, but does not change in the other. We show that these distribution-level patterns result from differences in the relative frequency of certain patterns of epistasis at the level of individual mutations, including fitness-correlated and idiosyncratic epistasis. 

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3. Patterns of Fitness and Gene Expression Epistasis Generated by Beneficial Mutations in the rho and rpoB Genes of Escherichia coli during High-Temperature Adaptation

   https://doi.org/10.1093/molbev/msae187  

   González-González, Andrea; Batarseh, Tiffany_N; Rodríguez-Verdugo, Alejandra; Gaut, Brandon_S; Samhita, ed., Laasya (September 2024, Molecular Biology and Evolution)

   Abstract Epistasis is caused by genetic interactions among mutations that affect fitness. To characterize properties and potential mechanisms of epistasis, we engineered eight double mutants that combined mutations from the rho and rpoB genes of Escherichia coli. The two genes encode essential functions for transcription, and the mutations in each gene were chosen because they were beneficial for adaptation to thermal stress (42.2 °C). The double mutants exhibited patterns of fitness epistasis that included diminishing returns epistasis at 42.2 °C, stronger diminishing returns between mutations with larger beneficial effects and both negative and positive (sign) epistasis across environments (20.0 °C and 37.0 °C). By assessing gene expression between single and double mutants, we detected hundreds of genes with gene expression epistasis. Previous work postulated that highly connected hub genes in coexpression networks have low epistasis, but we found the opposite: hub genes had high epistasis values in both coexpression and protein–protein interaction networks. We hypothesized that elevated epistasis in hub genes reflected that they were enriched for targets of Rho termination but that was not the case. Altogether, gene expression and coexpression analyses revealed that thermal adaptation occurred in modules, through modulation of ribonucleotide biosynthetic processes and ribosome assembly, the attenuation of expression in genes related to heat shock and stress responses, and with an overall trend toward restoring gene expression toward the unstressed state. 

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4. The roles of movement and coat proteins in the transport of tobamoviruses between plant cells

   https://doi.org/10.3389/fpls.2025.1580554  

   Kan, Yumin; Citovsky, Vitaly (April 2025, Frontiers in Plant Science)

   Tobamovirusis a large group of positive-sense, single-stranded RNA viruses that cause diseases in a broad range of plant species, including many agronomically important crops. The number of knownTobamovirusspecies has been on the rise in recent years, and currently, this genus includes 47 viruses. Tobamoviruses are transmitted mainly by mechanical contact, such as physical touching by hands or agricultural tools; and some are also transmitted on seeds, or through pollinator insects. The tobamoviral genome encodes proteins that have evolved to fulfill the main conceptual task of the viral infection cycle - the spread of the invading virus throughout the host plant cells, tissues, and organs. Here, we discuss this aspect of the infection cycle of tobamoviruses, focusing on the advances in our understanding of the local, i.e., cell-to-cell, and systemic, i.e., organ-to-organ, virus movement, and the viral and host plant determinants of these processes. Specifically, we spotlight two viral proteins—the movement protein (MP) and the coat protein (CP), which are directly involved in the local and systemic spread of tobamoviruses—with respect to their phylogeny, activities during viral movement, and interactions with the host determinants of the movement process. 

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5. Role of Viral Hemorrhagic Septicemia Virus Matrix (M) Protein in Suppressing Host Transcription

   https://doi.org/10.1128/JVI.00279-17  

   Ke, Qi; Weaver, Wade; Pore, Adam; Gorgoglione, Bartolomeo; Wildschutte, Julia Halo; Xiao, Peng; Shepherd, Brian S.; Spear, Allyn; Malathi, Krishnamurthy; Stepien, Carol A.; et al (July 2017, Journal of Virology)

   Viral Hemorrhagic Septicemia virus (VHSV) is a pathogenic fish rhabdovirus found in discrete locales throughout the northern hemisphere. VHSV infection of fish cells leads to upregulation of the host's virus detection response, but the virus quickly suppresses interferon (IFN) production and antiviral genes expression. By systematically screening each of the six VHSV structural and nonstructural genes, we have identified matrix protein (M) as its most potent anti-host protein. VHSV-IVb M alone suppressed mitochondrial antiviral signaling protein (MAVS) and type I IFN-induced gene expression in a dose-dependent manner. M also suppressed the constitutively active SV40 promoter and globally decreased cellular RNA levels. Chromatin immunoprecipitation (ChIP) studies illustrated that M inhibited RNA polymerase II (RNAP II) recruitment to gene promoters, and decreased RNAP II CTD Ser2 phosphorylation during VHSV infection. However, transcription directed by RNAP I-III was suppressed by M. To identify regions of functional importance, M proteins from a variety of VHSV strains were tested in cell-based transcriptional inhibition assays. M protein of a particular VHSV-Ia strain, F1, was significantly less potent than -IVb M at inhibiting SV40/luc expression, yet differed by just four amino acids. Mutation of D62 to alanine alone, or in combination with an E181 to alanine mutation (D62A/E181A), dramatically reduced the ability of -IVb M to suppress host transcription. Introducing either M D62A or D62A/E181A mutations into VHSV-IVb via reverse genetics resulted in viruses that replicated efficiently but exhibited less cytotoxicity and reduced anti-transcriptional activities, implicating M as a primary regulator of cytopathicity and host transcriptional suppression. Importance: Viruses must suppress host antiviral responses to replicate and spread between hosts. In these studies, we identified the matrix protein of the deadly fish Novirhabdovirus, VHSV, as a critical mediator of host suppression during infection. Our studies indicated that M alone could block cellular gene expression at very low expression levels. We identified several subtle mutations in M that were less potent at suppressing host transcription. When these mutations were engineered back into recombinant viruses, the resulting viruses replicated well but elicited less toxicity in infected cells and activated host innate immune responses more robustly. These data demonstrated that VHSV M plays an important role in mediating both virus-induced cell toxicity and viral replication. Our data suggest that its roles in these two processes can be separated to design effective attenuated viruses for vaccine candidates. 

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