Recombinant Lactobacillus johnsonii Segregation and condensation protein B (scpB)

What is the role of Segregation and condensation protein B (scpB) in Lactobacillus johnsonii chromosome dynamics?

The scpB protein plays a crucial role in chromosome organization and segregation during cell division in L. johnsonii. Based on research in Bacillus subtilis, disruption of scpB leads to temperature-sensitive slow growth, defects in chromosome structure, and formation of anucleate cells . ScpB forms a complex with ScpA and SMC (Structural Maintenance of Chromosomes) proteins that co-localize to discrete foci associated with DNA, typically located adjacent to chromosomal origin regions .

For researchers investigating this protein, it's important to note that the ScpB protein is part of a prokaryotic condensation complex that functions similarly to eukaryotic condensins. The complex forms condensation "factories" that organize DNA within the cell, pulling DNA away from mid-cell into both cell halves . This function is particularly important in L. johnsonii, which must maintain genomic integrity while surviving in challenging gastrointestinal environments.

How does scpB interact with scpA and SMC proteins to form a functional complex?

The interaction between scpB, scpA, and SMC forms a functional complex essential for proper chromosome dynamics. Research using the FRET technique and immunoprecipitation assays has demonstrated that ScpA and ScpB are directly associated with each other and with SMC in vivo . The complex operates with:

• SMC proteins forming a hinge-like structure to embrace DNA

• ScpA acting as a linker protein, connecting SMC to ScpB

• ScpB enhancing the stability of the complex and regulating its activity

These proteins co-localize to specific foci that are dynamically positioned based on the cell cycle stage - typically at mid-cell in young cells, and within both cell halves in older cells . All three components (SMC, ScpA, and ScpB) are necessary for the formation of these foci and proper chromosome organization. Genes similar to scpA and scpB are present in many bacteria and archaea, suggesting their products form a condensation complex with SMC in most prokaryotes .

What expression systems are most effective for producing recombinant L. johnsonii scpB?

For optimal expression of recombinant L. johnsonii scpB, researchers should consider:

Bacterial Expression Systems:

• E. coli-based systems with inducible promoters (T7, tac)

• Lactobacillus-based expression systems when studying interactions with other L. johnsonii proteins

Expression Optimization Strategies:

• Codon optimization based on the host's codon usage preferences

• Addition of affinity tags (His6, GST) for purification

• Temperature optimization during induction (lower temperatures often improve proper folding)

• Media selection (animal derivative-free media like TIL has shown better efficiency compared to conventional MRS media for some Lactobacillus proteins)

Expression Conditions Table:

When working with potentially membrane-associated proteins like ScpB, consider testing both cytoplasmic expression and periplasmic targeting to determine optimal localization for proper folding.

What purification methods yield the highest purity of recombinant L. johnsonii scpB?

To achieve high purity of recombinant L. johnsonii scpB, a multi-step purification strategy is recommended:

Initial Capture:

• Immobilized Metal Affinity Chromatography (IMAC) for His-tagged scpB

• Glutathione affinity chromatography for GST-fusion proteins

Intermediate Purification:

• Ion exchange chromatography based on scpB's theoretical isoelectric point

• Hydrophobic interaction chromatography for separating contaminants with different hydrophobicity

Polishing Step:

• Size exclusion chromatography to remove aggregates and achieve >85% purity

Quality Control:

• SDS-PAGE to assess purity (aim for >85% as achieved with recombinant scpA)

• Western blotting with anti-His or specific antibodies

• Mass spectrometry to confirm protein identity and integrity

• Functional assays to verify activity

For researchers working with L. johnsonii proteins, it's important to note that removal of endotoxins is crucial if the protein will be used in immunological studies, as L. johnsonii has been shown to interact with immune cells .

What methodologies are most effective for studying the in vivo localization of scpB in L. johnsonii cells?

For investigating the in vivo localization of scpB in L. johnsonii, researchers should consider these methodological approaches:

Fluorescent Protein Fusion Strategy:

• Construct C-terminal and N-terminal fusions with fluorescent proteins (YFP, CFP)

• Express under native promoters for physiological levels

• Validate functionality through complementation tests in scpB deletion strains

• Employ similar approaches to those used successfully with B. subtilis, where fusions of ScpA and ScpB to YFP or CFP revealed co-localization to discrete foci

Advanced Microscopy Techniques:

• Time-lapse fluorescence microscopy to track dynamic localization during cell cycle

• Super-resolution microscopy (STORM, PALM) for detailed sub-cellular positioning

• FRET analysis to study protein-protein interactions in vivo, as successfully demonstrated with ScpA and ScpB in B. subtilis

Data Analysis Methods:

• Quantitative image analysis to measure protein concentration and movement

• Co-localization analysis with DNA (DAPI staining) and origin markers

• 3D reconstruction to understand spatial organization

Researchers should expect to observe discrete foci that are present at mid-cell in young cells and within both cell halves, generally adjacent to chromosomal origin regions, in older cells - similar to the pattern observed in B. subtilis . The foci should be dependent on the presence of both SMC and ScpA proteins.

How do single amino acid substitutions in scpB affect complex formation with scpA and SMC?

Investigating the effects of single amino acid substitutions in scpB on complex formation requires a systematic approach:

Strategic Mutation Design:

• Identify conserved residues through multiple sequence alignment of scpB homologs

• Target predicted interface regions based on structural models

• Create a panel of alanine substitutions at conserved positions

• Design charge-reversal mutations at potential electrostatic interaction sites

Complex Formation Analysis Methods:

• In vivo FRET analysis to quantify changes in protein-protein interactions

• Co-immunoprecipitation assays to assess complex integrity

• Bacterial two-hybrid screening to identify critical interaction residues

• Surface plasmon resonance (SPR) with purified components to measure binding kinetics

Functional Impact Assessment:

• Monitor localization patterns of fluorescently-tagged mutant proteins

• Quantify the formation of discrete foci in cells

• Assess chromosome segregation efficiency and anucleate cell formation

Based on research with B. subtilis, mutations that disrupt the ScpA-ScpB interaction would be expected to show similar phenotypes to scpB deletion, including temperature-sensitive growth and defective chromosome segregation . The epistatic relationship between scpB, scpA, and smc genes suggests that mutations affecting complex formation would result in similar phenotypic consequences as complete gene disruption.

How does temperature affect the functionality and structural stability of recombinant L. johnsonii scpB?

The temperature-dependent functionality of scpB provides a valuable experimental system for researchers:

Temperature Effects on ScpB Function:
Based on studies in B. subtilis, disruption of scpB leads to temperature-sensitive growth defects above 23°C . This suggests that:

• ScpB function may be intrinsically temperature-sensitive

• The SMC-ScpA-ScpB complex integrity might be thermolabile

• Temperature-dependent changes in DNA topology may increase reliance on ScpB function

Experimental Approaches for Assessing Temperature Effects:

Biochemical Analysis Methods:

• Circular dichroism spectroscopy to monitor secondary structure changes with temperature

• Differential scanning calorimetry to determine melting temperatures

• Size exclusion chromatography at different temperatures to assess oligomerization state

• Thermal shift assays to identify stabilizing buffer conditions

For researchers working with recombinant L. johnsonii scpB, it's critical to control temperature precisely during purification and functional studies, as even small temperature fluctuations may significantly impact protein behavior and experimental outcomes.

What approaches can reveal the transcriptional regulation of scpB expression in L. johnsonii under different growth conditions?

Understanding the transcriptional regulation of scpB requires comprehensive genomic and molecular approaches:

Promoter Analysis and Regulatory Element Identification:

• 5' RACE to precisely map transcription start sites

• Reporter gene fusions (luciferase, GFP) to quantify promoter activity

• DNase footprinting and electrophoretic mobility shift assays to identify protein-DNA interactions

• ChIP-seq to identify transcription factors binding to the scpB promoter in vivo

Transcriptional Response Analysis:

• qRT-PCR to measure scpB expression under various conditions

• RNA-seq to place scpB regulation in the context of global transcriptional networks

• Single-cell RNA-seq to investigate cell-to-cell variability in expression

Environmental Factors to Test:

• Temperature shifts (particularly important given temperature-sensitive phenotypes)

• Growth phase dependence (log vs. stationary)

• Bile exposure (shown to affect L. johnsonii gene expression)

• Nutritional status (carbon, nitrogen limitation)

• pH stress (relevant to gastrointestinal survival)

Of particular interest would be examining whether scpB expression changes in response to bile, as bile has been shown to promote L. johnsonii extracellular vesicle production through transcriptional changes . Whole transcriptome analysis of L. johnsonii N6.2 grown with bile revealed upregulation of several peptidoglycan hydrolases and genes involved in fatty acid utilization , suggesting potential co-regulation with cell envelope remodeling processes.

How can in vitro reconstitution systems be developed to study the molecular mechanism of L. johnsonii ScpB in chromosome condensation?

Developing in vitro reconstitution systems for studying L. johnsonii ScpB function requires systematic biochemical approaches:

Protein Component Preparation:

• Express and purify full-length SMC, ScpA, and ScpB proteins with appropriate tags

• Prepare fluorescently labeled versions for FRET and single-molecule studies

• Generate domain constructs to identify minimal functional units

• Ensure proper folding through circular dichroism and activity assays

DNA Substrate Preparation:

• Circular DNA mimicking bacterial chromosomes

• Linear DNA fragments with specific sequences or structures

• Fluorescently labeled DNA for visualization

• Tethered DNA constructs for single-molecule manipulation

Assay Development Table:

Advanced Biophysical Approaches:

• Single-molecule FRET to observe conformational changes

• Total internal reflection fluorescence (TIRF) microscopy to visualize DNA-protein complexes

• Optical tweezers to measure forces during DNA compaction

• Cryo-electron microscopy to determine complex structure at different functional states

By reconstituting the SMC-ScpA-ScpB complex with DNA in vitro, researchers can directly examine how these proteins work together to organize and compact DNA, providing mechanistic insights that complement in vivo observations of chromosome segregation and condensation.

What role does scpB play in L. johnsonii's adaptation to the host gastrointestinal environment?

Investigating scpB's role in L. johnsonii's adaptation to the gastrointestinal environment requires integrating molecular genetics with host-microbe interaction studies:

Stress Response Analysis:

• Create scpB conditional mutants and reporter strains

• Expose to relevant GI stressors:

  • Bile (particularly important as bile promotes L. johnsonii vesiculation)

  • Acid stress (pH transitions in GI tract)

  • Oxidative stress (immune cell-derived ROS)

  • Nutrient limitation and competition

Bile Response Mechanisms:
Bile exposure significantly impacts L. johnsonii physiology, with transcriptome analysis revealing upregulation of cell wall modifying enzymes and fatty acid utilization genes . These changes suggest extensive cell envelope remodeling that may require coordinated chromosome organization by the ScpB-containing condensin complex.

Host Colonization Studies:

• Compare wild-type and scpB mutant colonization efficiency in animal models

• Analyze competitive fitness using mixed infections

• Examine persistence during antibiotic treatment or immune challenge

• Assess spatial distribution along the GI tract

Immune Interaction Characterization:
L. johnsonii interacts with host immune cells and can modulate dendritic cell function . Researchers should investigate whether ScpB indirectly influences these interactions by:

• Affecting surface protein display through chromosome organization

• Influencing stress responses that alter immunomodulatory functions

• Enabling adaptation to changing host conditions

By connecting chromosome dynamics (mediated by ScpB) to stress adaptation mechanisms, researchers can gain insights into how fundamental cellular processes contribute to L. johnsonii's success as a probiotic microorganism in the challenging gastrointestinal environment.

What are the optimal protocols for creating precise gene deletions or mutations in scpB in L. johnsonii?

Creating precise genetic modifications in L. johnsonii requires specialized approaches due to the challenges of working with lactic acid bacteria:

CRISPR-Cas9 Based Genome Editing:

• Design sgRNAs targeting scpB with minimal off-target effects

• Prepare repair templates with desired mutations and homology arms (800-1000bp)

• Use temperature-sensitive plasmids for transient CRISPR-Cas9 expression

• Screen transformants using PCR and sequencing verification

Traditional Homologous Recombination Approach:

• Design suicide vectors containing homology regions flanking scpB

• Include counter-selectable markers (sacB, upp) for enrichment of double crossover events

• Optimize transformation protocols with glycine treatment to weaken cell walls

• Confirm gene replacements using both PCR and Southern blotting

Creating Conditional Mutants:
For essential genes or to study temperature-sensitive phenotypes similar to those observed in B. subtilis scpB mutants :

• Implement xylose-inducible promoter systems similar to those used in B. subtilis studies

• Develop degron-tag approaches for controlled protein depletion

• Create temperature-sensitive alleles by targeted mutagenesis

Verification Methods Table:

For researchers working with L. johnsonii, it's crucial to optimize transformation efficiency by adjusting cell wall weakening treatments and electroporation parameters, as transformation can be challenging in Lactobacillus species.

How can the interaction between recombinant L. johnsonii scpB and DNA be characterized biochemically?

Characterizing the DNA-binding properties of recombinant L. johnsonii scpB requires multiple complementary biochemical techniques:

DNA Binding Assays:

• Electrophoretic Mobility Shift Assays (EMSA)

  • Use radiolabeled or fluorescently labeled DNA fragments

  • Test sequence-specific vs. non-specific binding

  • Examine effects of ScpA and SMC on binding characteristics

• Fluorescence Anisotropy

  • Measures binding affinity (Kd) in solution

  • Allows real-time binding kinetics assessment

  • Can be performed under various buffer conditions

• Surface Plasmon Resonance (SPR)

  • Provides association and dissociation rate constants

  • Enables study of complex formation dynamics

  • Allows testing of different DNA structures

DNA Structure Preference Analysis:

• Compare binding to linear vs. supercoiled DNA

• Test preferential binding to bent DNA or specific structures

• Examine effects of DNA topology on binding

Footprinting and Crosslinking:

• DNase I footprinting to identify protected regions

• Hydroxyl radical footprinting for high-resolution contacts

• Photo-crosslinking to capture transient interactions

• ChIP-seq or related techniques for genome-wide binding profiles

Based on studies of the SMC-ScpA-ScpB complex, researchers should examine both direct DNA binding by ScpB alone and how ScpB modulates the DNA interaction properties of the entire condensin complex. The observed foci formation at specific chromosomal locations in vivo suggests potential preferential binding to certain DNA regions or structures that should be investigated in biochemical assays.

What proteomics approaches can reveal post-translational modifications of L. johnsonii scpB?

Comprehensive characterization of post-translational modifications (PTMs) on L. johnsonii scpB requires multi-faceted proteomics approaches:

Sample Preparation Strategies:

• Enrich for ScpB using affinity purification from:

  • Recombinant expression systems

  • Native L. johnsonii under various growth conditions

  • Different subcellular fractions

• Employ multiple digestion strategies:

  • Trypsin (standard)

  • Chymotrypsin (complementary coverage)

  • Glu-C (alternative cleavage patterns)

Mass Spectrometry Methods:

• Shotgun proteomics for initial PTM discovery

• Targeted approaches for verification of specific modifications

• Top-down proteomics for intact protein analysis

• Electron transfer dissociation (ETD) for labile modifications

PTMs to Investigate:

Quantitative Analysis:

• SILAC or TMT labeling to compare modification levels under different conditions

• Parallel reaction monitoring for accurate quantification

• Label-free approaches for relative abundance estimation

Researchers should pay particular attention to how PTMs might change in response to environmental conditions relevant to L. johnsonii's lifestyle, such as bile exposure , acid stress, or temperature shifts , which could reveal regulatory mechanisms controlling ScpB function during adaptation to the gastrointestinal environment.

How can computational modeling predict the structural features of L. johnsonii scpB?

Computational modeling of L. johnsonii scpB structure requires a multi-level approach that integrates various prediction methods:

Sequence-Based Structure Prediction:

• Secondary structure prediction using PSIPRED, JPred

• Disorder prediction with PONDR, IUPred

• Domain identification using InterPro, Pfam

• Coiled-coil region prediction with COILS, Marcoil

Homology Modeling Pipeline:

• Template identification using HHpred, BLAST

• Alignment optimization with structural considerations

• Model building with MODELLER, SWISS-MODEL

• Refinement using molecular dynamics simulations

• Validation with PROCHECK, QMEAN

Advanced Modeling Approaches:

• AlphaFold2 or RoseTTAFold for deep learning-based prediction

• Integrative modeling combining experimental data (SAXS, crosslinking)

• Molecular dynamics simulations to assess conformational dynamics

• Protein-protein docking to predict interactions with ScpA and SMC

Function Prediction:

• Binding site prediction using CASTp, COACH

• Electrostatic surface analysis to identify potential DNA interaction regions

• Conservation mapping to highlight functionally important residues

• Normal mode analysis to predict flexible regions and conformational changes

Based on studies of homologous proteins, researchers should expect L. johnsonii ScpB to contain predominantly α-helical secondary structures and coiled-coil regions , which are common features in proteins involved in chromosome dynamics. The model should also focus on interface regions that might mediate interaction with ScpA, as these proteins have been shown to associate directly .

How can single-cell proteomics be applied to study scpB dynamics in L. johnsonii populations?

Single-cell proteomics offers powerful approaches to understand cell-to-cell variation in scpB expression and function:

Sample Preparation for Single-Cell Analysis:

• Microfluidic isolation of individual L. johnsonii cells

• Nanodroplet processing for protein extraction

• Miniaturized digestion protocols

• Label or label-free quantification strategies

Mass Spectrometry Approaches:

• Targeted proteomics (SRM/MRM) focusing on ScpB and related proteins

• Data-independent acquisition for broader coverage

• Carrier proteome approaches to boost sensitivity

• SCoPE2 methodology for increased throughput

Data Analysis Strategies:

• Specialized computational pipelines for single-cell data

• Normalization methods accounting for technical variation

• Statistical approaches for sparse data

• Machine learning for pattern recognition

Biological Questions to Address:

• Cell cycle-dependent changes in ScpB abundance

• Correlation between ScpB levels and chromosome segregation efficiency

• Population heterogeneity in response to environmental stressors

• Co-expression patterns with ScpA and SMC

Single-cell proteomic data can be integrated into the Single-cell Proteomic DataBase (SPDB) , which provides visualization tools and analysis pipelines specifically designed for this type of data. This integration would allow researchers to compare ScpB dynamics across different experimental conditions and in relation to other proteins involved in chromosome organization.

What are the optimal experimental designs for studying the temperature-sensitive phenotypes of scpB mutants?

Designing experiments to investigate temperature-sensitive phenotypes of scpB mutants requires careful consideration of multiple factors:

Temperature Control Strategy:

• Use precise temperature-controlled incubators (±0.1°C precision)

• Implement gradient temperature experiments (23-37°C range)

• Design temperature shift protocols (permissive to restrictive)

• Include recovery experiments (restrictive to permissive)

Experimental Design Table:

Controls and Considerations:

• Include wild-type controls at all temperatures

• Test complemented strains (native scpB expression)

• Assess effects on ScpA and SMC localization

• Monitor for suppressor mutations (whole genome sequencing)

Data Analysis Approaches:

• Quantitative image analysis for nucleoid morphology

• Statistical methods for comparing growth parameters

• Mathematical modeling of temperature dependence

• Single-cell tracking for lineage analysis

Based on research in B. subtilis, researchers should focus on temperatures above 23°C, where scpB disruption leads to growth defects and chromosome segregation problems . The temperature-sensitive phenotype provides an excellent tool for studying the function of this protein complex, allowing for conditional inactivation and observation of immediate consequences.

How can multi-omics approaches be integrated to understand the role of scpB in L. johnsonii physiology?

Integrating multi-omics approaches provides a comprehensive understanding of scpB's role in L. johnsonii:

Multi-omics Data Collection:

• Genomics: Whole genome sequencing of wild-type and mutant strains

• Transcriptomics: RNA-seq under various conditions

• Proteomics: Global protein expression and PTMs

• Metabolomics: Metabolic profiles and fluxes

• Phenomics: Growth characteristics and stress responses

Integration Strategies:

• Network analysis to identify co-regulated genes and proteins

• Pathway enrichment to understand functional implications

• Correlation analysis across different data types

• Machine learning for pattern recognition

Biological Context Focus:

• Cell cycle regulation and chromosome dynamics

• Stress response pathways (particularly bile and temperature stress)

• Host-microbe interaction networks

• Probiotic and immunomodulatory functions

Data Visualization and Analysis Platforms:

• Specialized tools for multi-omics integration

• Network visualization software

• Custom R or Python pipelines for cross-platform analysis

• Database submission to relevant repositories (SPDB for proteomic data)

A particularly interesting approach would be to examine how bile exposure, which significantly affects L. johnsonii gene expression and physiology , influences ScpB function and the entire chromosome organization machinery. This could reveal connections between environmental adaptation and fundamental cellular processes like chromosome segregation.

What statistical approaches should be used to analyze localization patterns of fluorescently tagged scpB in L. johnsonii?

Analyzing localization patterns of fluorescently tagged scpB requires robust statistical methods:

Image Analysis Pipeline:

• Image preprocessing (background subtraction, deconvolution)

• Spot detection and segmentation

• Feature extraction (intensity, size, shape)

• Cell cycle stage classification

• Spatial distribution analysis

Statistical Analysis Methods:

• Bayesian approaches for spot detection reliability

• Kernel density estimation for spatial distribution patterns

• Bootstrapping for confidence interval determination

• Mixed-effects models to account for cell-to-cell variability

Quantitative Metrics Table:

Advanced Analytical Approaches:

• Machine learning classification of localization patterns

• Hidden Markov Models for temporal dynamics

• Spatial statistics (Ripley's K function, nearest neighbor analysis)

• Correlation analysis with chromosome markers

Based on studies in B. subtilis, researchers should expect to observe dynamic patterns with foci present at mid-cell in young cells, and within both cell halves, generally adjacent to chromosomal origin regions, in older cells . Statistical comparison between wild-type and mutant strains can reveal how specific mutations affect these localization patterns and their correlation with proper chromosome segregation.

How can structural knowledge of L. johnsonii scpB inform rational protein engineering for enhanced function?

Rational engineering of L. johnsonii scpB requires detailed structural understanding and systematic modification strategies:

Structure-Based Engineering Approaches:

• Stability enhancement through:

  • Disulfide bond introduction at strategic positions

  • Surface charge optimization

  • Hydrophobic core redesign

• Interaction interface optimization:

  • Enhancing ScpA binding affinity

  • Modifying DNA interaction properties

  • Tuning SMC complex dynamics

• Domain fusion and protein chimeras:

  • Creating ScpA-ScpB fusion proteins for enhanced complex formation

  • Generating chimeric proteins with domains from thermophilic organisms for increased stability

  • Developing split-protein complementation systems for monitoring interactions

Functional Enhancement Strategies Table:

Validation Methods:

• In vitro biochemical assays with purified engineered proteins

• Growth complementation studies in scpB mutant strains

• Microscopy to assess chromosomal organization

• Competition assays to measure fitness effects

By understanding the structural basis of ScpB function in the SMC-ScpA-ScpB complex, researchers can design improved variants with enhanced stability, controlled activity, or novel functions that could provide insights into chromosome dynamics or lead to strains with improved characteristics for probiotic applications.

What are the implications of scpB research for developing improved L. johnsonii strains for probiotic applications?

Understanding scpB function has significant implications for probiotic development:

Strain Improvement Strategies:

• Enhanced stress tolerance through:

  • Optimized chromosome organization under stress conditions

  • Improved bile resistance by tuning scpB expression or activity

  • Temperature resilience through engineered scpB variants

• Increased genetic stability:

  • Reduced anucleate cell formation during manufacturing

  • Decreased mutation rates through optimized DNA organization

  • Prevention of plasmid loss in recombinant strains

• Improved host interaction:

  • Enhanced survival in gastrointestinal conditions

  • Optimized growth and colonization properties

  • Potential effects on immunomodulatory functions

Probiotic Performance Metrics:

• Survival during processing and storage

• Resistance to gastric acid and bile

• Colonization efficiency

• Therapeutic efficacy in disease models

• Safety profile and antibiotic resistance patterns

Research Direction Priorities:

• Connect chromosome biology to stress adaptation mechanisms

• Investigate how scpB function influences L. johnsonii's probiotic properties

• Develop strains with optimized scpB expression for specific applications

• Explore interactions between chromosome organization and metabolite production

Research has shown that L. johnsonii has significant therapeutic potential in various diseases and that growing conditions affecting cellular physiology (like bile exposure ) can influence its beneficial properties. Understanding how fundamental processes like chromosome segregation contribute to these characteristics could lead to rationally designed probiotic strains with enhanced efficacy.

How might the study of L. johnsonii scpB contribute to our broader understanding of bacterial chromosome biology?

L. johnsonii scpB research offers unique insights into bacterial chromosome biology:

Comparative Evolutionary Perspectives:

• Analysis of ScpB across diverse bacteria reveals:

  • Conserved mechanisms in chromosome organization

  • Species-specific adaptations in condensin complexes

  • Evolution of prokaryotic chromosome management systems

• Comparison with eukaryotic systems:

  • Functional parallels between prokaryotic and eukaryotic condensins

  • Divergent strategies for similar challenges

  • Evolutionary origins of chromosome organization mechanisms

Fundamental Questions Addressed:

• How bacteria organize chromosomes without a nuclear membrane

• Mechanisms of DNA compaction in different bacterial species

• Coordination between chromosome organization and cell division

• Environmental adaptation of chromosome dynamics

Technology Development Opportunities:

• New tools for visualizing chromosome organization

• Novel antimicrobial targets based on chromosome segregation

• Synthetic biology applications of engineered chromosome organization

Cross-disciplinary Connections:

• Physical principles of DNA organization in confined spaces

• Computational modeling of chromosome dynamics

• Systems biology of genome organization networks

The epistatic relationship between scpB, scpA and smc genes suggests that these proteins form a functional module conserved across diverse bacteria. By comparing the specific properties of L. johnsonii ScpB with homologs from other species, researchers can identify both universal principles and specialized adaptations in bacterial chromosome biology.