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Record W2045019620 · doi:10.1074/jbc.m114.607481

Structural and Functional Characterization of a Ketosteroid Transcriptional Regulator of Mycobacterium tuberculosis

2014· article· en· W2045019620 on OpenAlex

Why this work is in the frame

A frame that forgets how it found something cannot be audited. These are the routes that admitted this work.

affAt least one author lists a Canadian institution in the pinned OpenAlex snapshot.
fundA Canadian funder is recorded on the work.

Bibliographic record

VenueJournal of Biological Chemistry · 2014
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicSteroid Chemistry and Biochemistry
Canadian institutionsUniversity of British ColumbiaUniversity of Toronto
FundersArgonne National LaboratoryBiological and Environmental ResearchNational Institute of General Medical SciencesCanadian Institutes of Health ResearchNational Institutes of HealthMichael Smith Health Research BCU.S. Department of Energy
KeywordsIsothermal titration calorimetryChemistryStereochemistryDimerLigand (biochemistry)RepressorBiochemistryGeneGene expressionReceptor

Abstract

fetched live from OpenAlex

Background: KstR2 regulates cholesterol catabolic genes in Mycobacterium tuberculosis. Results: Dimeric KstR2 Mtb binds two molecules of HIP-CoA with high affinity. Each binding site spans the subunits and includes residues conserved in TetR family repressors (TFRs) that bind CoA thioesters. Conclusion: HIP-CoA binding to KstR2 Mtb induces a conformation that abrogates DNA binding. Significance: The study identifies molecular determinants of cholesterol catabolism and CoA binding in TFRs. Catabolism of host cholesterol is critical to the virulence of Mycobacterium tuberculosis and is a potential target for novel therapeutics. KstR2, a TetR family repressor (TFR), regulates the expression of 15 genes encoding enzymes that catabolize the last half of the cholesterol molecule, represented by 3a-H-4(3-propanoate)-7a-methylhexahydro-1,5-indane-dione (HIP). Binding of KstR2 to its operator sequences is relieved upon binding of HIP-CoA. A 1.6- resolution crystal structure of the KstR2 Mtb HIP-CoA complex reveals that the KstR2 Mtb dimer accommodates two molecules of HIP-CoA. Each ligand binds in an elongated cleft spanning the dimerization interface such that the HIP and CoA moieties interact with different KstR2 Mtb protomers. In isothermal titration calorimetry studies, the dimer bound 2 eq of HIP-CoA with high affinity (K d 80 10 nM) but bound neither HIP nor CoASH. Substitution of Arg-162 or Trp-166, residues that interact, respectively, with the diphosphate and HIP moieties of HIP-CoA, dramatically decreased the affinity of KstR2 Mtb for HIP-CoA but not for its operator sequence. The variant of R162M that decreased the affinity for HIP-CoA (G 13 kJ mol 1 ) is consistent with the loss of three hydrogen bonds as indicated in the structural data. A 24-bp operator sequence bound two dimers of KstR2. Structural comparisons with a ligand-free rhodococcal homologue and a DNA-bound homologue suggest that HIP-CoA induces conformational changes of the DNA-binding domains of the dimer that preclude their proper positioning in the major groove of DNA. The results provide insight into KstR2-mediated regula-tion of expression of steroid catabolic genes and the determinants of ligand binding in TFRs.

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

Full frame distilled prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.

metaresearch head score (Codex)0.000
metaresearch head score (Gemma)0.000
Version: codex-gemma-dda1882f352aValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.062
Threshold uncertainty score0.539

Codex and Gemma teacher scores by category

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0000.000
Research integrity0.0000.000
Insufficient payload (model declined to judge)0.0000.000

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.011
GPT teacher head0.206
Teacher spread0.195 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it