TB9.4 Family assigned · low

H37Rv Rv3208A · MTBC0 - · 90 aa · 3585677–3585949 (-) · RefSeq YP_177943.1

Annotation: from legacy to revised

Legacy (H37Rv / Mycobrowser)hypothetical protein
MTBC0 PGAP re-annotation
Revised (this work)Structurally characterized DUF3107 protein (Pfam) of unknown biochemical function. RefSeq leaves it 'hypothetical protein'. A crystal structure of the hexadecameric form of this family was solved (PDB 7rur; Foldseek prob 1.0, E 3e-11, TM 0.95). STRING context links it to the cytochrome bcc subunit qcrC (Rv2194). Family/structure assigned; molecular function unfixed.

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 Q6MWZ8 TrEMBL · unreviewed · Evidence at protein level
UniProt nameConserved protein TB9.4

UniProt still lists this protein as Conserved protein TB9.4; the revised annotation above is ahead of the current UniProt record.

Functional vocabulary (eggNOG-mapper, orthology transfer)

COG category S Function unknown
eggNOG descriptionProtein of unknown function (DUF3107)
Orthologous group2E3M2

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 0.0 · strong purifying
Polymorphic sites (≥ 0.1% of strains) 2 synonymous, 0 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
DUF3107PF11305.14 3.6e-311–73 Protein of unknown function (DUF3107)

Structural neighbours (Foldseek on the ESMFold model, exploratory)

ESMFold model confidence: mean pLDDT 69.8 (low). Low-confidence model: the fold may be unreliable, so treat these structural hits with caution.

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
7rur-assembly1_A 1.00 0.93 3.5e-10 sig 7rur-assembly1_A Crystal structure of the hexadecameric form of Rv3208A
7rur-assembly1_B 1.00 0.93 3.5e-10 sig 7rur-assembly1_B Crystal structure of the hexadecameric form of Rv3208A
7rus-assembly1_A 1.00 0.90 2.0e-10 sig 7rus-assembly1_A Crystal structure of the octameric form of Rv3208A
7rur-assembly1_C 1.00 0.92 3.7e-10 sig 7rur-assembly1_C Crystal structure of the hexadecameric form of Rv3208A
2rd1-assembly2_B 0.97 0.60 8.6e-02 2rd1-assembly2_B X-Ray structure of the protein Q7CQI7. Northeast Structural Genomics Consortium target StR87A
1zq1-assembly1_B 0.96 0.52 4.1e-02 1zq1-assembly1_B Structure of GatDE tRNA-Dependent Amidotransferase from Pyrococcus abyssi
3hfo-assembly1_B 0.94 0.55 1.1e-01 3hfo-assembly1_B Crystal Structure of an Hfq protein from Synechocystis sp.
2rb6-assembly3_B-2 0.91 0.58 1.0e-01 2rb6-assembly3_B-2 X-Ray structure of the protein Q8EI81. Northeast Structural Genomics Consortium target SoR78A

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

Closest characterised functional partner: qcrC (ubiquinol-cytochrome C reductase cytochrome subunit C), high confidence from genomic context alone (score 717 excluding text-mining). This association is the citable seed of a function hypothesis for this hypothetical protein.

PartnerProductScoreNo text-miningChannels (≥400)
Rv2194 qcrC ubiquinol-cytochrome C reductase cytochrome subunit C 716 717 ctx cooccurence:714
Rv3209 hyp hypothetical protein 705 706 ctx neighborhood:704
Rv2199c ctaF cytochrome c oxidase polypeptide 4 675 676 ctx cooccurence:662
Rv2195 qcrA ubiquinol-cytochrome C reductase rieske iron-sulfur subunit 613 614 ctx cooccurence:608
Rv2169c transmembrane protein 514 515 ctx cooccurence:513
Rv2698 transmembrane protein 511 511 ctx cooccurence:504
Rv2196 qcrB ubiquinol-cytochrome C reductase cytochrome subunit B 496 497 ctx cooccurence:482
Rv3013 hyp hypothetical protein 487 488 ctx cooccurence:482
Rv1828 HTH-type transcriptional regulator 482 482 ctx cooccurence:479
Rv0528 transmembrane protein 453 454 ctx cooccurence:451
Rv0004 hyp hypothetical protein 449 449 ctx cooccurence:449
Rv2680 hyp hypothetical protein 446 447 ctx cooccurence:444
Rv2229c hyp hypothetical protein 445 445 ctx cooccurence:442
Rv1234 transmembrane protein 428 428 ctx cooccurence:423
Rv2219A membrane protein 408 409

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

  • Pfam DUF3107
  • Foldseek -> 7rur (hexadecameric Rv3208 family), prob 1.0, E 3e-11, TM 0.95
  • STRING anchor qcrC/Rv2194 (respiratory chain)
  • Structural homology: AlphaFold DB model + Foldseek vs PDB (project 'Still unknown gene function', phase5-7, 2026-06-10). A fold-level family assignment, not a demonstrated activity.

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_177943.1)
  • Domains: Pfam-A via hmmscan --cut_ga — DUF3107 (PF11305.14)
  • 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 2E3M2
  • Curated reference: UniProt Q6MWZ8 (TrEMBL, unreviewed; Evidence at protein level)
  • 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 69.8, low)
  • Interaction network: STRING v12.0 (Szklarczyk et al. 2023, doi:10.1093/nar/gkac1000), taxon 83332, CC-BY 4.0 — 15 functional partner(s); context anchor qcrC
  • Primary literature: none located yet; annotation rests on the domain/homology sources above.

Ancestral MTBC0 protein sequence

>H37Rv|Rv3208A|TB9.4
MEVKIGITDSPRELVFSSAQTPSEVEELVSNALRDDSGLLTLTDERGRRFLIHTARIAYVEIGVADARRVGFGVGVDAAAGSAGKVATSG