Rv3294c Still unknown · low

H37Rv Rv3294c · MTBC0 - · 269 aa · 3675186–3675995 (-) · RefSeq YP_177954.1

Annotation: from legacy to revised

Legacy (H37Rv / Mycobrowser)hypothetical protein
MTBC0 PGAP re-annotation
Revised (this work)Conserved hypothetical; function unknown. Structure-based hint rejected: mismatch endonuclease NucS/EndoMS is Rv1321; this gene is a DUF4143 protein (paralogue-of-fold).

Annotated on the H37Rv protein: this gene has no 1:1 ancestral MTBC0 anchor (PE/PPE, paralogue, IS element, or otherwise unanchored CDS).

Curated reference (UniProt)

UniProt L7N658 TrEMBL · unreviewed · Predicted
UniProt nameDUF4143 domain-containing protein

Functional vocabulary (eggNOG-mapper, orthology transfer)

COG category J Translation, ribosomal structure and biogenesis
eggNOG descriptionDomain of unknown function (DUF4143)
Orthologous groupCOG1373
KEGG orthology K07133

Orthology-based transfer (eggNOG 5.0.2, diamond). EC/KO/GO/CAZy are computed annotations, not manual curation; cross-check against the primary literature before treating a specific reaction as established.

Conservation & selection (intra-MTBC, 145 209 strains)

pN/pS 1.116 · relaxed/neutral
Polymorphic sites (≥ 0.1% of strains) 1 synonymous, 3 missense, 0 nonsense, 0 frameshift

pN/pS from segregating SNPs (singletons removed) normalised by possible sites. Low pN/pS = purifying selection (a strong signal that a "hypothetical" is a real, constrained gene). A high pN/pS is ambiguous: relaxed constraint or positive selection (drug resistance, antigenic variation) inflate it; e.g. rpoB/katG/pncA score high here for resistance, not loss of function. A clonal disruption (one allele over a clade) suggests lineage pseudogenisation; a convergent one (many independent alleles) is typical of resistance loss-of-function.

Domains (Pfam, hmmscan --cut_ga)

PfamAccessioni-EvalueResiduesDescription
DUF4143PF13635.13 1.8e-4153–214 Domain of unknown function (DUF4143)

Structural neighbours (Foldseek on the ESMFold model, exploratory)

ESMFold model confidence: mean pLDDT 87.0 (confident). A confident model makes the fold comparison meaningful.

Best matches against the PDB, ranked by Foldseek homology probability. A high probability / TM-score suggests a shared fold; unless flagged sig (E < 0.01) these are fold hypotheses, not assignments.

TargetProbTME-valueDescription
2dk5-assembly1_A 0.99 0.71 5.4e-02 2dk5-assembly1_A Solution structure of Winged-Helix domain in RNA polymerase III 39KDa polypeptide
5gkh-assembly1_B 0.97 0.55 2.1e-02 5gkh-assembly1_B Structure of EndoMS-dsDNA2 complex
1r1u-assembly1_B 0.97 0.55 3.0e-02 1r1u-assembly1_B Crystal structure of the metal-sensing transcriptional repressor CzrA from Staphylococcus aureus in the apo-form
5f6f-assembly1_B 0.97 0.45 2.4e-02 5f6f-assembly1_B S. aureus MepR G34R Mutant
5fb2-assembly1_A 0.96 0.41 1.3e-02 5fb2-assembly1_A S. aureus MepR F27L Mutant bound to oligodeoxyribonucleotide
5ffx-assembly2_D 0.95 0.43 2.2e-02 5ffx-assembly2_D S. aureus MepR G34K Mutant
5ffz-assembly1_A-2 0.95 0.46 3.8e-02 5ffz-assembly1_A-2 S. aureus MepR bound to ethidium bromide
2vld-assembly1_B 0.93 0.50 1.7e-02 2vld-assembly1_B crystal structure of a repair endonuclease from Pyrococcus abyssi

Functional interaction network (STRING v12, guilt-by-association)

Closest characterised functional partner: Rv3295 (TetR family transcriptional regulator), medium confidence from genomic context alone (score 609 excluding text-mining). This association is the citable seed of a function hypothesis for this hypothetical protein.

PartnerProductScoreNo text-miningChannels (≥400)
Rv2810c Probable transposase; Rv2810c, (MTCY16B7.33), len: 133 aa. Probable transposase for IS1555, similar to C-terminal domain of transposases for 733 733 coexpression:733
Rv3295 TetR family transcriptional regulator 610 609 ctx neighborhood:605
Rv0962c lprP lipoprotein LprP 553 553 coexpression:553
Rv3297 nei endonuclease VIII 493 494 ctx neighborhood:491
Rv3296 lhr ATP-dependent helicase 469 469 ctx neighborhood:464
Rv0919 GCN5-like N-acetyltransferase 443 443 ctx cooccurence:436

STRING combines evidence channels (neighborhood, fusion, cooccurrence, coexpression, experimental, database, text-mining) into a 0–1000 score. The ctx badge marks edges carried by the genomic-context channels (conserved neighborhood, fusion, phylogenetic co-occurrence), which are independent of orthology and structure and the strongest signal for an unknown gene. The no text-mining column recomputes the score from data alone, so a link that does not depend on the literature is visible. Association is a function hypothesis, not proof: corroborate with the operon context and the primary literature before assigning a function.

Evidence

  • Structural Foldseek hit not propagated -- mismatch endonuclease NucS/EndoMS is Rv1321; this gene is a DUF4143 protein (paralogue-of-fold)
  • Reviewed against literature (extended structural cross-check, 2026-06-02)

Sources

  • Ancestral sequence & coordinates: Harrison LB et al. (2024), An imputed ancestral reference genome for the MTBC, doi:10.1101/2023.09.07.556366
  • Product annotation: NCBI PGAP on MTBC0; legacy from H37Rv NC_000962.3 (RefSeq YP_177954.1)
  • Domains: Pfam-A via hmmscan --cut_ga — DUF4143 (PF13635.13)
  • Sequence-level signal: ESM Atlas (EvolutionaryScale × BioHub) — exploratory
  • Controlled vocabulary: eggNOG-mapper 2.1.12 (Cantalapiedra et al. 2021, doi:10.1093/molbev/msab293), eggNOG 5.0 DB (Huerta-Cepas et al. 2019) — OG COG1373
  • Curated reference: UniProt L7N658 (TrEMBL, unreviewed; Predicted)
  • Intra-MTBC selection: pN/pS and disruption from SPDI variants of 145 209 MTBC strains (this work, local collection vs H37Rv NC_000962.3)
  • Model confidence: ESMFold per-residue pLDDT (mean 87.0, confident)
  • Interaction network: STRING v12.0 (Szklarczyk et al. 2023, doi:10.1093/nar/gkac1000), taxon 83332, CC-BY 4.0 — 6 functional partner(s); context anchor Rv3295
  • Primary literature: none located yet; annotation rests on the domain/homology sources above.

Ancestral MTBC0 protein sequence

>H37Rv|Rv3294c|
MGLPRRPCCDTTGSARYRESVRRYPRIGEDSAAYRRRLCRESAKARNVDRVVKRDAADVSNLQRIADLPRLIRLLAARSASELNLSSLATDAEIPVRTLPPYLDLLETLYLIDRIPAWSTNLSKRVVDRPKVLLLDSGLAARLVNVSPTGAGPHANPNAAGAIIETFVIAELRRQLGWSQQAPRLFHYRDRDGAEVDLILETADGLIAAIEIKSAATLRGRDTRSISRLRDKVGARFAGGVILHTGPQAQPFGDRLAAVPIDILWSPSG