Biology subjects

Anand, R.

Publications and source records attributed to Anand, R..

At least 19 recordsLinked to original sources

bioRxiv2026

Hierarchical Temporal Transformer for Cancer Grade Prediction and Cross-Cancer Transfer Learning from Pathology Reports

Background: Language models for cancer reports usually read one document at a time and are tested on tumour types seen in training. Continuous-time sequence models address the first gap, but whether they use visit timing, not just order, is seldom tested. Methods: We developed the Hierarchical Temporal Transformer (HTT), which encodes reports with adapted BiomedBERT and reads the sequence with a cancer-type-conditioned temporal transformer. We evaluated it against best-possible AUROC ceilings on two synthetic benchmarks, one with the label stated in the text and one recoverable only from tumour sizes and visit intervals (ten seeds). Diagnostics led to an interval-aware time code (HTT-IA). On 4,786 TCGA pathology reports in 14 types, three types were withheld, and a text-only HTT was compared with a survival model. Results: On the phrase benchmark HTT reached AUROC 0.931 +/- 0.012 against 0.913 +/- 0.007 for a last-report baseline (+0.018, 95% CI -0.009 to +0.050); dates cannot raise the ceiling there. On the growth benchmark the initial time code scored 0.790, equal to a no-time model (0.787), whereas HTT-IA reached 0.843 (+0.053, 0.026 to 0.082) against ceilings of 0.894 without and 0.979 with dates. On unseen tumour types HTT reached 0.925 +/- 0.007, equal to in-distribution performance and to a single-report baseline. Text-only macro C-index was 0.592 (CALM 0.626-0.652). Conclusion: Cross-type transfer is carried by the pre-trained encoder. Visit timing is used only when the data make it identifiable and the time code is on a clinical scale. Ceilings, order-only ablations and re-dating probes are inexpensive checks for such models.

bioinformatics↗

bioRxiv2026

Layered phage therapy exploits capsule-loss escape and altered antibiotic susceptibility in CG23 hypervirulent Klebsiella pneumoniae

The rise of drug-resistant Klebsiella pneumoniae infections is prompting a re-evaluation of phage therapy as an alternative treatment. Phage therapy has achieved successful outcomes, yet important limitations persist, including the rapid emergence of phage resistance and the time required to source suitable phages. Combining multiple phages into a phage cocktail based on genetic diversity, morphology or distinct infection mechanisms is a common strategy to mitigate resistance. Phage pairing based on these criteria neither mitigates nor delays the emergence of resistance, using CG23 hypervirulent Klebsiella pneumoniae (hvKp) EC0675 as a model. In CG23 Kp strains, resistance arises through disruption of the capsular biosynthetic pathway, leading to capsule impairment and global cross-resistance to all capsule-active phages. Combining phages active against encapsulated wild-type bacteria with those active against capsule-deficient escape mutants can overcome this constraint, improving bacterial suppression in vitro and reducing bacterial burdens across multiple organs in a mouse systemic infection model. In parallel, phage-driven capsule loss increased susceptibility of hvKp EC0675 to selected antibiotics targeting the bacterial envelope or cell wall in vitro. These findings highlight the contribution of capsule-associated resistance to phage cross-resistance in the CG23 Kp strains examined, and support further evaluation of phage combinations targeting both wild-type bacteria and resistant derivatives, including their use alongside selected antibiotics, informing future clinical translation.

microbiology↗

bioRxiv2026

Assessment of antimicrobial potential of cell-free supernatant from Lacticaseibacillus paracasei SRG isolated from the vaginal microbiota of healthy women against uropathogens

Among women, urinary tract infection (UTI) represents a major health concern. Approximately 50% of the adult female population experienced an episode of UTI at least once. UTI and recurrent UTIs caused by resistant bacteria are very common in clinical settings. The treatment and management of UTI is becoming more challenging because of the rise in antibiotic-resistant bacteria. Lactobacillus spp are the dominant bacterial genus in urinary and vaginal microbiota, which are also used as probiotics. In this study, Lacticaseibacillus paracasei SRG isolated from the vagina of healthy women were evaluated for its probiotic-associated properties and cell-free supernatant (CFS)-mediated antibacterial activity against eight uropathogens. The antibacterial activity of CFS was concentration- and pH-dependent, which was significantly evident at CFS/2 and CFS/4 concentrations and acidic pH values of 4 and 5. The antibacterial activity in CFS remains unaltered even after heat and proteinase K treatment, which suggests the role of heat- and protease-resistant metabolites beside the contributions from acidic conditions. L. paracasei SRG showed strong autoaggregation (85.32%) and cell-surface hydrophobicity (55.60%). In addition, it also exhibits co-aggregation with all indicator strains, suggesting substantial interaction with diverse pathogenic bacteria. These observations highlighted the L. paracasei SRG as a potential probiotic candidate developed from indigenous vaginal microbiota.

microbiology↗

bioRxiv2026

TRF2 mediates force-resistant bridging of distal telomeric DNA repeats

Shelterin binds telomeres, regulates telomere length, and protects chromosome ends from mis-repair, which is essential for genome stability. Telomere end-protection uniquely depends on the TRF2 subunit, which assembles telomeric DNA into a protective T-loop structure in vivo, yet the biochemical mechanism underlying this process remains unresolved. Using single-molecule optical tweezers and atomic force microscopy, we show that the human shelterin complex tethers distal telomeric DNA repeats, looping the intervening DNA. These bridges withstand tens of piconewtons of applied load, rupture at a force that scales with telomeric repeat number and re-form on relaxation. Structure-function analysis demonstrates that this force-resistant tethering is TRF2-dependent under our conditions and depends on multiple interfaces, with the TRFH domain making a major but non-exclusive contribution: introducing the TRF2 TRFH domain into TRF1 is sufficient to confer tethering, whereas its deletion from TRF2 reduces but does not abolish activity. We propose telomeric dsDNA tethering by TRF2 as a candidate first step in T-loop formation, preceding invasion of the telomeric 3' ssDNA overhang.

biophysics↗

bioRxiv2026

Hierarchical temporal transformer for cancer grade prediction and cross cancer transfer learning from pathology reports

Language models for cancer clinical reports carry two blind spots. They read each report in isolation, ignoring how a patients disease changes across visits, and they are evaluated only on cancer types present in their training data. We present the Hierarchical Temporal Transformer (HTT), a two-level architecture that addresses both. Level 1 encodes each report with BiomedBERT adapted by low-rank adaptation (LoRA). Level 2 is a temporal transformer that reads a patients full report sequence using a continuous-time positional encoding built from the measured number of days between visits, with learnable cancer-type embeddings supplying per-family conditioning. Two experiments test the two capabilities separately, since no fully open corpus contains longitudinal reports for many cancer types. On a controlled synthetic corpus of sequential radiology reports, in which progression phrases are inserted from templated trajectories, HTT reaches a validation AUROC of 0.942 against 0.881 for a single-report baseline and transfers to held-out pancreatic cancer at 0.995 against 0.949 while the two models are indistinguishable on a 60-patient test set. On 4,786 real pathology reports from the TCGA-Reports corpus spanning 14 cancer types HTT predicts tumor grade for three types withheld entirely from training, reaching AUROC 1.000 on thyroid carcinoma, 0.960 on sarcoma and 0.808 on lung squamous cell carcinoma. The mean held-out AUROC of 0.923 equals the in-distribution test AUROC of 0.923, so transfer to unseen cancer families incurred no measurable penalty. Ablation on the real corpus shows that the transfer is carried by the pre-trained encoder rather than by the temporal components, which, with one report per patient, contribute 0.39 AUROC points. Grade-related pathological language therefore appears to be learnable in a cancer-type agnostic way, which points toward unified cancer NLP systems that require no per-type retraining.

bioinformatics↗

bioRxiv2026

Genomic hallmarks of parasexual reproduction in three hybrid groups of the human pathogen Cryptococcus neoformans

Hybridization is a major driver of fungal evolution, yet knowledge of the molecular mechanisms underpinning hybridization and its genomic impact remain limited. Here, we analyse 197 Cryptococcus neoformans genomic sequences, including 13 newly sequenced strains, identifying three genetically clustered and distinct hybrid groups (H1, H2 and H3) each with unique parental origins and ecological associations. Using phylogenomics, population structure analyses, and long-read genome assemblies, we identified hybrid genomes with chromosome-wide loss of heterozygosity (LOH), inheritance of large intact parental haplotype blocks and widespread aneuploidy within and across genomes. These patterns are also observed when progeny were generated with spo11{Delta} parents, indicating these features are a result of a meiotic-independent process such as parasex. This is further underscored by discovery of haploid and near-haploid recombinants in both spo11{Delta} mutant progeny and reanalysed wild-type hybrid progeny. We hypothesise these haploid and near-haploid recombinants are generated through ploidy reduction via independent chromosome assortment because of concerted chromosomal loss, which is a key feature of the parasexual cycle. Phenotypic assays demonstrated that several hybrid isolates have diverse growth and virulence patterns, underscoring functional consequences of genome plasticity. Together, our work suggests a non-meiotic reproductive process contributes to shape the genotypic and phenotypic diversity of Cryptococcus neoformans.

genomics↗

bioRxiv2026

A comparative analysis of urinary microbiome identifies putative probiotics

Urinary tract infections (UTIs) are the most prevalent bacterial infections globally, and their management increasingly challenged by antimicrobial resistance (AMR). Probiotics offer a promising approach to mitigate AMR by competitively excluding uropathogens and enhancing host immunity by producing immune modulators. Despite being potential, key gaps persist between the discovery of uroprotective probiotic strains and optimization of formulations for urinary tract delivery. Here, we analyzed the urinary microbiome of UTI patients and healthy individuals to identify potential probiotic candidates for the prevention and management of UTIs. Publicly available 16S rRNA amplicon sequencing data of the urinary tract were processed using a standardized pipeline for sequence quality assessment, taxonomic assignment, and microbial function prediction. Comparative analysis showed a significant shift in microbial composition between UTI patients and healthy controls. The dominated phyla identified included Acidobacteriota, Actinobacteriota, Bacteroidota, Campylobacterota, Cyanobacteria, Firmicutes, Fusobacteriota, Patescibacteria, Proteobacteria, and Synergistota. Overall differential abundance analysis revealed Escherichia coli as the predominant UTI-associated species, while Lactobacillus crispatus was enriched in healthy samples. Additionally, predictive functional analysis indicated that metabolic pathways associated with beneficial microbes were enriched in the healthy group. Overall, the study highlights the association of distinct urinary microbiome signatures with infection status, which supports L. crispatus as the most promising probiotic for UTI prevention and control.

bioinformatics↗

bioRxiv2026

Role of a childhood cancer-linked BRIP1/FANCJ germline variant in genomic instability and cancer cell vulnerability

Childhood cancer is frequently associated with inherited pathogenic variants of cancer predisposition genes. Using Whole-Exome Sequencing, we identified an inherited, monoallelic pediatric cancer-linked germline variant of DNA helicase FANCJ/BRIP1, BRIP1R162Q of unclear clinical significance. Intriguingly, in vitro helicase assays demonstrated that BRIP1R162Q encodes a hyperactive DNA helicase. In cells, stable expression of BRIP1R162Q in BRIP1-proficient cells selectively sensitized to ectopic DNA replication stress. Moreover, BRIP1R162Q expressing cells showed chronic, steady-state activation of DNA replication stress indicated by decreased fork speed, fork stalling and impaired fork symmetry. Moreover, BRIP1R162Q expression triggered genomic instability indicated by elevated {gamma}H2AX foci numbers and chromosomal aberrations, suggesting BRIP1R162Q as a variant driving genomic instability. Mechanistically, BRIP1R162Q mislocalizes upon replication stress, and leads to accumulation of DNA secondary G-quadruplex (G4) structures and R-loops. RNaseH1 expression, which resolves R-loops, also reduced G4 levels and relieved BRIP1R162Q-induced replication stress. Finally, BRIP1R162Q introduces exploitable vulnerabilities for targeted therapies using G4-ligand Pyridostatin and DNA damage kinase ATR and DNA-PK inhibition. Our findings propose a mechanistic framework of how a childhood cancer-linked monoallic, hypermorphic BRIP1 germline variant participates in cancer development, and suggest potential therapeutic treatment strategies.

cancer biology↗

bioRxiv2026

MIC13-linked cristae disruption causes metabolic failure and early fibrotic remodelling in mitochondrial liver disease

Mitochondrial diseases are highly complex and heterogeneous, and nearly 20% of cases involve severe liver pathology. Here, we report an affected individual carrying a pathogenic MIC13 variant (c.260-2A>G) associated with early-onset mitochondrial hepato-encephalopathy. Such mitochondrial hepatopathies are rare, multisystemic disorders with major liver involvement, difficult to diagnose, lack effective treatment, and are poorly understood, in part due to the absence of faithful disease-relevant cellular models. To investigate hepatocyte-specific consequences of the MIC13 variant, we generated iPSCs carrying this disease-causing variant and differentiated them into induced hepatocytes (iHeps). MIC13, a key component of the MICOS complex required for cristae formation, was disrupted in these cells, and the resultant iHeps exhibited the same cristae defects observed in clinical samples. Integrated multi-omics and biochemical analyses revealed extensive metabolic rewiring, including disrupted amino acid turnover and accumulation of tricarboxylic acid (TCA) and urea cycle intermediates. Additionally, profound alterations in methionine cycle and transsulfuration pathways, along with enhanced bile acid synthesis, collectively affect methylation potential, redox homeostasis, and detoxification. Lipid metabolism was also impaired, with incomplete {beta}-oxidation, increased ketogenesis, and diminished lipid storage. At the cellular level, extensive extracellular matrix (ECM) remodelling, increased intracellular collagen accumulation and enhanced cell migration indicated an early fibrotic phenotype. Overall, this clinically relevant model uncovers mechanistically how cristae defects drive metabolic imbalance and hepatocyte dysfunction, ultimately leading to early fibrotic changes in mitochondrial liver disease. These findings provide a strong mechanistic foundation for understanding mitochondrial liver disease and developing targeted therapeutic strategies.

biochemistry↗

bioRxiv2025

Gabapentinoids promote striatal dopamine release and rescue multiple deficits of a mouse model of early Parkinson's

Neuronal entry and handling of intracellular calcium have long-been hypothesised to burden vulnerable dopamine neurons in Parkinsons disease. However, no treatments for Parkinsons target calcium biology. Gabapentinoid drugs bind to 2{delta} subunits of voltage-gated calcium channels (VGCCs) and are licensed for neurological disorders including dopamine-dysregulated restless leg syndrome, suggesting their potential utility to modify both calcium biology and dopamine signalling. We therefore tested whether gabapentinoids modulate dopamine signalling, underlying VGCC-dependence, and potential for treating Parkinsons. In mouse striatum, we reveal that gabapentinoids ex vivo promote dopamine release, via sex-specific dependence on 2{delta}1/2 subunits and alterations to the calcium- and VGCC-subtype-dependence of dopamine release or its tonic inhibition by striatal GABA. In vivo administration of gabapentinoids to a mouse model of early Parkinsons rescued deficits in dopamine release, dysregulation of GABAergic inhibition and dopamine content, and abolished parkinsonian deficits in movement transitions. Thus, gabapentinoids urgently deserve attention for repurposing for Parkinsons disease. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=184 SRC="FIGDIR/small/675586v1_ufig1.gif" ALT="Figure 1"> View larger version (51K): org.highwire.dtl.DTLVardef@16cd7e1org.highwire.dtl.DTLVardef@d104e5org.highwire.dtl.DTLVardef@98683eorg.highwire.dtl.DTLVardef@1e3e3ae_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

bioRxiv2023

Mitochondrial Apolipoprotein MIC26 is a metabolic rheostat regulating central cellular fuel pathways

Mitochondria play central roles in metabolism and metabolic disorders such as type 2 diabetes. MIC26, a MICOS complex subunit, was linked to diabetes and modulation of lipid metabolism. Yet, the functional role of MIC26 in regulating metabolism under hyperglycemia is not understood. We employed a multi-omics approach combined with functional assays using WT and MIC26 KO cells cultured in normoglycemia or hyperglycemia, mimicking altered nutrient availability. We show that MIC26 has an inhibitory role in glycolysis and cholesterol/lipid metabolism under normoglycemic conditions. Under hyperglycemia, this inhibitory role is reversed demonstrating that MIC26 is critical for metabolic adaptations. This is partially mediated by alterations of mitochondrial metabolite transporters. Furthermore, MIC26 deletion led to a major metabolic rewiring of glutamine utilization as well as oxidative phosphorylation. We propose that MIC26 acts as a metabolic rheostat, that modulates mitochondrial metabolite exchange via regulating mitochondrial cristae, allowing cells to cope with nutrient overload.

cell biology↗

bioRxiv2023

SLP2 coordinates MICOS assembly and cristae morphogenesis via MIC13 and YME1L

The MICOS complex subunit MIC13 is essential for mitochondrial cristae organization. Mutations in MIC13 cause severe mitochondrial hepato-encephalopathy displaying defective cristae morphology and loss of the MIC10-subcomplex. Here we identified stomatin-like protein 2 (SLP2) as an interacting partner of MIC13 and decipher a critical role of SLP2 as an auxiliary MICOS subunit, modulating cristae morphology. SLP2 provides a large interaction hub for MICOS subunits and loss of SLP2 leads to drastic alterations in cristae morphology. Double deletion of SLP2 and MIC13 showed reduced assembly of core MICOS subunit, MIC60 into MICOS and dispersion of MIC60-specific puncta, demonstrating a critical role of SLP2-MIC13 in MICOS assembly and crista junction (CJ) formation. We further identified that the mitochondrial i-AAA protease YME1L in coordination either with MIC13 or SLP2 differentially regulates MICOS assembly pathways thereby interlinking MIC13-specific or scaffolding-specific role of SLP2 with quality control and assembly of the MICOS complex. YME1L- depletion in MIC13 KO could restore MIC10-subcomplex and reform the nascent CJ. Taken together, we propose seeder model for MICOS assembly and CJ formation, where SLP2- MIC13 seed the assembly of MIC60 into MICOS complex and promote the formation of CJ by regulating the quality and stability of MIC10-subcomplex.

biochemistry↗

bioRxiv2023

Identifying Selectivity Filters in Protein Biosensor for Ligand Screening

Specialized sensing mechanisms in bacteria enable the identification of cognate ligands with remarkable selectivity in highly xenobiotic-polluted environments, where these ligands are utilized as energy sources. Here, via an integrated all-atom computer simulation, biochemical assay and isothermal titration calorimetry approaches we determine the molecular basis of MopR, a phenol biosensors complex selection process of ligand entry. Our results reveal a set of strategically placed selectivity filters along the ligand entry pathway of MopR. These filters act as checkpoints, screening diverse aromatic ligands at the protein surface based on their chemical features and sizes. Ligands meeting specific criteria are allowed to enter the sensing site in an orientation-dependent manner. Sequence and structural analyses demonstrate the conservation of this ligand entry mechanism across the sensor class, with individual amino acids along the selectivity filter path playing a critical role in ligand selection. Together, this investigation highlights the importance of interactions with the ligand entry pathway, in addition to interactions within the binding pocket, for achieving ligand selectivity in biological sensing. The findings enhance our understanding of ligand selectivity in bacterial phenol biosensors and provide insights for the rational expansion of the biosensor repertoire, particularly for the biotechnologically relevant class of aromatic pollutants.

biochemistry↗

bioRxiv2023

Live-cell super-resolution nanoscopy reveals modulation of cristae dynamics in bioenergetically compromised mitochondria

Cristae membranes have been recently shown to undergo intramitochondrial merging and splitting events. Yet, the metabolic and bioenergetic factors regulating them are unclear. Here we investigated whether and how cristae membrane remodelling is dependent on oxidative phosphorylation (OXPHOS) complexes, the mitochondrial membrane potential ({Delta}{Psi}m), and the ADP/ATP nucleotide translocator. Advanced live-cell STED nanoscopy combined with in-depth quantification were employed to analyse cristae morphology and dynamics after treatment of mammalian cells with rotenone, antimycin A, oligomycin A and CCCP. This led to formation of enlarged mitochondria along with reduced cristae density but did not change the number of cristae remodelling events. CCCP treatment leading to {Delta}{Psi}m abrogation even enhanced the cristae dynamics showing their {Delta}{Psi}m-independent nature. Inhibition of OXPHOS complexes was accompanied by reduced ATP levels but did not affect cristae dynamics. However, inhibition of ADP/ATP exchange led to aberrant cristae morphology and impaired cristae dynamics in a mitochondrial subset. In sum, we provide quantitative data of cristae membrane remodelling under different conditions supporting an important interplay between OXPHOS, metabolite exchange and cristae membrane dynamics. Summary BlurbCristae morphology and dynamics are intricately connected

cell biology↗

bioRxiv2022

The APE2 nuclease is essential for DNA double strand break repairby microhomology-mediated end-joining.

Microhomology-mediated end-joining (MMEJ) is an intrinsically mutagenic pathway of DNA double strand break repair essential for proliferation of homologous recombination (HR) deficient tumors. While targeting MMEJ has emerged as a powerful strategy to eliminate HR-deficient (HRD) cancers, this is limited by an incomplete understanding of the mechanism and factors required for MMEJ repair. Here, we identify the APE2 nuclease as a novel MMEJ effector. We show that loss of APE2 blocks the fusion of deprotected telomeres by MMEJ and inhibits MMEJ in DNA repair reporter assays to levels comparable to Pol Theta suppression. Mechanistically, we demonstrate that APE2 possesses intrinsic flap-cleaving activity, that its MMEJ function in cells depends on its nuclease domain and further identify uncharacterized domains required for recruitment to damaged DNA. We conclude that HR-deficient cells are addicted to APE2 due to a previously unappreciated role in MMEJ, which could be exploited in the treatment of cancer.

molecular biology↗

bioRxiv2022

Phenol Sensing in Nature Modulated via a Conformational Switch Governed by Dynamic Allostery

NtrC family of proteins sense external stimuli and accordingly stimulate stress and virulence pathways via activation of associated {sigma}54-dependent RNA polymerases. Here, we establish that MopR, an NtrC protein, harbors a dynamic bi-directional electrostatic network that connects the phenol pocket to two distal regions, namely the "G-hinge" and the "allo-steric-linker". While G-hinge influences the entry of phenol, the allosteric-linker passes the signal to the downstream ATPase domain. Phenol binding induces a rewiring of the electrostatic connections by eliciting dynamic allostery, and it was demonstrated that perturbation of the core relay residues results in a complete loss of ATPase stimulation. A mutation of the G-hinge,[~]20[A] from the phenol pocket, demonstrated altered flexibility by shifting the pattern of conformational states accessed, leading to a protein with 7-fold enhanced phenol binding ability and enhanced transcriptional activation. A global analysis illustrates that dynamic allostery-driven conserved community networks are universal and evolutionarily conserved across species.

biochemistry↗

bioRxiv2022

Alteration of myocardial structure and function in RAF1associated Noonan syndrome: Insights from cardiac disease modeling based on patient-derived iPSCs

Noonan syndrome (NS), the most common among the RASopathies, is caused by germline variants in genes encoding components of the RAS-MAPK pathway. Distinct variants, including the recurrent Ser257Leu substitution in RAF1, are associated with severe hypertrophic cardiomyopathy (HCM). Here, we investigated the elusive mechanistic link between NS-associated RAF1S257L and HCM using three-dimensional cardiac bodies and bioartificial cardiac tissues generated from patient-derived induced pluripotent stem cells (iPSCs) harboring the pathogenic RAF1 c.770C>T missense change. We characterize the molecular, structural and functional consequences of aberrant RAF1 -associated signaling on the cardiac models. Ultrastructural assessment of the sarcomere revealed a shortening of the I-bands along the Z disc area in both iPSC-derived RAF1S257L cardiomyocytes, and myocardial tissue biopsies. The disease phenotype was partly reverted by using both MEK inhibition, and a gene-corrected isogenic RAF1L257S cell line. Collectively, our findings uncovered a direct link between a RASopathy gene variant and the abnormal sarcomere structure resulting in a cardiac dysfunction that remarkably recapitulates the human disease. These insights represent a basis to develop future targeted therapeutic approaches.

cell biology↗

bioRxiv2021

Decoding the Mechanism of Specific RNA Targeting by Ribosomal Methytransferases

Methylation of specific nucleotides is integral for ribosomal biogenesis and serves as a common way to confer antibiotic resistance by pathogenic bacteria. Here, by determining the high-resolution structure of 30S-KsgA by cryo-EM, a state was captured, where KsgA juxtaposes between helices h44 and h45, separating them, thereby enabling remodeling of the surrounded rRNA and allowing the cognate site to enter the methylation pocket. With the structure as a guide, factors that direct the enzyme to its cognate site with high fidelity were unearthed by creating several mutant versions of the ribosomes, where interacting bases in the catalytic helix h45 and surrounding helices h44, h24, and h27 were mutated and evaluated for their methylation efficiency. The biochemical studies delineated specificity hotspots that enable KsgA to achieve an induced fit. This study enables the identification of distal exclusive allosteric pocket and other divergent structural elements in each rMTase, which can be exploited to develop strategies to reverse methylation, mediated drug resistance.

biochemistry↗