eccC3 Family assigned · medium auto-curated

Annotation: from legacy to revised

Auto-curated: this verdict and function were generated by rules from PGAP + Pfam + Foldseek and have not been hand-reviewed.

In the literature (TB corpus sweep) studied as much outside M. tuberculosis

6 TB publications mention this gene. 6 publications discuss this gene (count verified against title and abstract text). **Its biology is documented at least as much OUTSIDE M. tuberculosis as within it**: queried separately, 5 records carry a non-TB mycobacterial context (species named in the 5-record verified sample: M. abscessus (2), M. smegmatis (2), M. leprae (1)) against 3 with a M. tuberculosis context. The context counts come from distinct PubMed queries (the total is text-verified, the context counts are raw query volumes), so they are not parts of the total above and need not add up to it. Mycobacterial genetics is largely done in M. smegmatis, so part of what is 'known' about this gene is known by proxy. reformulation, not a quotation

IMPORTANT — 'better studied elsewhere' does NOT mean 'function established in M. tuberculosis'. Findings obtained in M. smegmatis (a non-pathogenic, fast-growing species with a different lifestyle and regulation), or in M. marinum / M. leprae / M. abscessus, do NOT transfer automatically to M. tuberculosis. Treat this body of work as CONTEXT to verify, not as settled knowledge. This distinguishes a gene that is dark because nobody has looked from one that is dark despite having been studied. Source: PubMed (whole): H37Rv locus tag + GENE NAME + ortholog identifiers (Mb…, MMAR_…, MSMEG_…, ML…, MAB_…), under a mycobacterial context filter; hits verified against the abstract text. Species-context counts distinguish M. tuberculosis literature from literature on other mycobacteria. phase76/phase77, 2026-07-13.

Genomic-neighbour overlap (structural caveat) co-directional · 0 % of gene

co-directional overlap: ordinary (e.g. shared stop/start codons in an operon), not the Rv2438A-type artefact P20.1, derived from GFF3 gene coordinates, 2026-08-03.

Conditional expression context (iModulons)

Member of 2 independently-modulated gene set(s): IdeR (ideR), Zur (zur).

iModulon membership (independently-modulated gene sets from a 647-sample RNA-seq compendium): the conditional co-expression context. Co-expression is a regulatory context, NOT a molecular function. Source: iModulonDB / modulome_mtb (Yoo 2022).

CRISPRi vulnerability

Vulnerability index -6.31 (95% CI -6.78 to -5.84). A more negative index = more vulnerable to knockdown (better drug-target quality); indicative threshold VI ≤ -6 = highly vulnerable.

Quantitative CRISPRi knockdown, graded (finer than binary Tn-seq essentiality). Source: CRISPRi vulnerability index (Bosch 2021, supplementary Table S2 'mmc3.xlsx', bulk import via Europe PMC -- supersedes the per-gene pebble.rockefeller.edu scrape of phase46_crispri_vulnerability.py).

Orthologues (reciprocal best hits across mycobacteria)

Reciprocal-best-hit orthologues (DIAMOND) against the Mycobrowser reference proteomes. A missing species is informative: e.g. a gene absent from M. leprae was likely lost in its reductive genome evolution. Locus tags link to Mycobrowser.

Curated reference (UniProt)

Functional vocabulary (eggNOG-mapper, orthology transfer)

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, 142 601 strains, snapshot of 2026-09-29)

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.

Outgroup conservation (beyond the MTBC) Actinomycetia

Two orthogonal outgroup signals. M. canettii (the immediate outgroup) gives a deep-divergence dN/dS (a low value confirms a constrained, real gene; shown as confident only at ≥8 substitutions, else flagged low-power). Genus-wide presence/absence (tblastn vs assembled non-MTBC genomes) places the gene on the ancient-core ↔ MTBC-specific axis: a gene absent even from the closest MTBAP relatives is a candidate MTBC-specific innovation (possible host-adaptation factor, to confirm by synteny). The phylostratum extends that axis outside the genus (tblastn vs 13 reference genomes spanning Mycobacteriaceae → Corynebacteriales → Actinomycetia → outside the phylum): it is the deepest clade in which a homolog is still detected, i.e. a proxy for gene age. Read it with the null model in mind: a shallow (young) stratum can also reflect homology-detection failure for short or fast-evolving ORFs, so it is a descriptive axis, not a proof of novelty.

Essentiality (transposon mutagenesis) essential

Genome-wide Himar1 transposon essentiality in H37Rv (DeJesus 2017). An essential call (ES/ESD/GD) is strong, independent evidence that a "hypothetical" locus encodes a functional, selectively required gene — orthogonal to intra-species conservation.

Chemical-genetic target & druggability (PROSPECT) hypomorph tool strain

This gene is part of the PROSPECT collection of TetON transcriptional-knockdown (hypomorph) strains of essential M. tuberculosis genes, built as a sensitised background for chemical-genetic mechanism-of-action deconvolution. Being in the panel means the gene is an essential / vulnerable target for which a validated knockdown tool strain exists.

Panel membership reflects essentiality/vulnerability and the availability of a genetic tool, not a specific molecular function; it never changes the verdict here. Source: Bond AN et al., Nat Commun 2025;16:9673 (doi:10.1038/s41467-025-64662-x); PROSPECT chemical-genetic platform.

Proteomics (mass spectrometry) detected

Detection by mass spectrometry is direct, experimental evidence that the protein product exists — orthogonal to sequence conservation and to Tn-seq essentiality, and especially decisive for a "hypothetical" locus. Read the dataset count as abundance, not as replication. Measured across this atlas (3,491 detected proteins), the number of datasets a protein appears in tracks its abundance almost monotonically: median 1.2 ppm at 1 dataset of 16, 27 ppm at 10, 392 ppm at 15, 1,325 ppm at 16 — three orders of magnitude. A high count therefore says the protein is abundant more than it says several independent experiments confirmed it. What the count does establish, at any value, is that the product exists.

Predicted localisation (DeepTMHMM + lipobox)

Transmembrane topology and signal peptide from DeepTMHMM (deep-learning reference predictor); lipoproteins from a (myco)bacterial lipobox motif. A sequence-based prediction of subcellular context.

Physico-chemical properties (computed, ProtParam)

Computed from the ancestral MTBC0 sequence with the ExPASy ProtParam method (Biopython). Descriptive biophysical context: a positive GRAVY flags a hydrophobic (often membrane) protein, a high instability index (>40) predicts a short in-vitro half-life, an extreme pI hints at compartment or binding partner.

Domains (Pfam, hmmscan --cut_ga)

3D structure (interactive — Mol*/pdbe-molstar)

Experimental structure 6j17 (PDBe), the strongest available structural evidence for this gene. Drag to rotate, scroll to zoom; hetero-groups (ligands, ions, waters hidden by default) shown as sticks.

Experimental structures (Protein Data Bank) 1 solved

Experimentally solved structures mapped from the UniProt accession via PDBe/SIFTS (1 total; up to 8 shown, ranked by sequence coverage then resolution). An experimental structure is direct proof of the folded product and the strongest structural evidence — superseding the predicted ESMFold/AlphaFold models below for any covered region.

Structural search (AlphaFold DB model, Foldseek vs PDB — genome-wide) pLDDT 84.6

Foldseek search of the AlphaFold DB model (mean pLDDT 84.6, gated at 70) against the PDB — a genome-wide extension of the ESMFold dark-gene search that also covers proteins beyond the single-sequence length limit. Confident structural neighbours (E < 0.01) shown.

Genomic context (neighbours & predicted operon) operon of 11

Neighbours from the H37Rv annotation (+ strand). The operon is predicted by co-directional intergenic distance (same strand, gaps ≤50 bp) — a transcription-unit hypothesis, not a mapped TSS. For a "hypothetical", co-transcription with a characterised operon is a concrete functional lead (complements the STRING neighborhood channel below).

Published operon evidence (two independent sources)

Two independent published operon maps, cited alongside the distance heuristic above: an association corroborating or contradicting the heuristic, never proof of shared function. REMap's RNA-seq coverage continuity is the most direct of the three signals; Roback's logistic regression and the intergenic-distance heuristic are statistical proxies.

Transcriptional regulation (signed TRN: ChIP-seq + TFOE)

Regulatory edges from the ISB signed transcriptional regulatory network (TF ChIP-seq binding, Minch 2015 + TF-overexpression response, Rustad 2014). An edge is regulatory evidence (binding and/or expression change), not necessarily direct. For a "hypothetical", membership in a known regulon (e.g. DosR dormancy, PhoP virulence) is a physiological-context lead, not a function.

Condition: Condition-blind by construction. These edges come from transcription-factor OVEREXPRESSION in a single laboratory condition, so a listed regulator is not a claim that it governs this gene in every circumstance. Measured counter-examples exist within this atlas (see Rv2516c, where the DosR regulon collapses under moxifloxacin while the gene itself rises). Read a regulator as 'capable of shifting this gene', and look for the condition before building a hypothesis on it.

Additional regulons (curated, outside the TFOE-derived TRN)

Curated from primary literature not covered by the ISB signed TRN (Minch 2015 / Rustad 2014): Zur regulon (Maciag et al. 2007, PMID 17098899), SigG regulon (Gaudion et al. 2013, PMID 23871545), LexA/SOS regulon (Smollett et al. 2012, PMID 22528497) — added P16.25 (2026-08-18); CRP regulon (Kahramanoglou et al. 2014, PMID 24957601) — added P16.25b (2026-09-27). Confidence and caveats are per-edge and carried from the source paper — read them before citing a specific gene as "regulated by" its listed TF. An entry reading "binding site only — direction not determined" is not a regulation claim: it records a measured ChIP-seq binding site for which the source reports no expression change, and the absence of such a change is not evidence of absent regulation.

Node colour = verdict, dashed = still uncharacterised; edge colour = evidence (green experimental, orange genomic-context, grey co-expression), width ∝ score. Click a partner to open its page; "Explore full network" to walk the graph.

Closest characterised functional partner: eccD3 (ESX-3 secretion system protein EccD), high confidence (score 1000, all STRING channels except text-mining; the partner is chosen on its genomic-context evidence). The pair's experimental or database support is transferred from orthologous pairs in other proteomes only: nothing was measured in M. tuberculosis, so it is an association inferred by orthology rather than measured in this organism.

Showing the 15 strongest of this gene's 51 partners at medium confidence (18 at high confidence). The table is an extract, not a neighbourhood: across the proteome the real median degree is 34 while the served median is 15, so no hub, degree distribution or centrality may be read on this table. None of the 51 partners rests on an interaction measured in M. tuberculosis itself.

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.

Where the experimental evidence comes from, and why it matters here. STRING's experimental channel fuses interactions measured in M. tuberculosis with interactions measured in another organism and transferred to this pair by orthology. The two are not equivalent for this atlas, whose standing rule is that better studied elsewhere is not a function established in M. tuberculosis. Measured on the served network: of the 38 229 partner pairs shown across the atlas, 13 483 partner rows carry a non-zero experimental channel, and only 106 pairs rest on an interaction measured in M. tuberculosis itself — 0.3 % of the network. Those 106 are long-established complexes (ESX-1/ESAT-6, RNA polymerase with CarD and RbpA, the Pup–Mpa proteasome, gyrase, Emb, GarA with PknB and PknG, the Rel modules); no still-uncharacterised gene is among them. The reason is not a gap in this atlas but in the literature: there is almost no experimental interactome of this bacterium, IntAct holding 134 curated protein pairs for a ~4 000-gene proteome. A exp (transferred) badge is therefore a lead, never a measurement, and an absence of badge is not evidence that the pair does not interact.

Evidence

• Legacy H37Rv annotation: ESX-3 secretion system protein EccC3
• MTBC0 PGAP product: type VII secretion system ESX-3 FtsK/SpoIIIE family ATPase EccC3
• Pfam (hmmscan --cut_ga): FtsK_SpoIIIE PF01580.25 (E=1e-58)
• (auto-curated by rules from PGAP + Pfam + Foldseek; not hand-reviewed)

Ancestral MTBC0 protein sequence

>mtbc0_000303|Rv0284|eccC3
MSRLIFEARRRLAPPSSHQGTIIIEAPPELPRVIPPSLLRRALPYLIGILIVGMIVALVATGMRVISPQTLFFPFVLLLAATALYRGNDKKMRTEEVDAERADYLRYLSVVRDNIRAQAAEQRASALWSHPDPTALASVPGSRRQWERDPHDPDFLVLRAGRHTVPLATTLRVNDTADEIDLEPVSHSALRSLLDTQRSIGDVPTGIDLTKVSRITVLGERAQVRAVLRAWIAQAVTWHDPTVLGVALAARDLEGRDWNWLKWLPHVDIPGRLDALGPARNLSTDPDELIALLGPVLADRPAFTGQPTDALRHLLIVVDDPDYDLGASPLAVGRAGVTVVHCSASAPHREQYSDPEKPILRVAHGAIERWQTGGWQPYIDAADQFSADEAAHLARRLSRWDSNPTHAGLRSAATRGASFTTLLGIEDASRLDVPALWAPRRRDEELRVPIGVTGTGEPLMFDLKDEAEGGMGPHGLMIGMTGSGKSQTLMSILLSLLTTHSAERLIVIYADFKGEAGADSFRDFPQVVAVISNMAEKKSLADRFADTLRGEVARREMLLREAGRKVQGSAFNSVLEYENAIAAGHSLPPIPTLFVVADEFTLMLADHPEYAELFDYVARKGRSFRIHILFASQTLDVGKIKDIDKNTAYRIGLKVASPSVSRQIIGVEDAYHIESGKEHKGVGFLVPAPGATPIRFRSTYVDGIYEPPQTAKAVVVQSVPEPKLFTAAAVEPDPGTVIADTDEQEPADPPRKLIATIGEQLARYGPRAPQLWLPPLDETIPLSAALARAGVGPRQWRWPLGEIDRPFEMRRDPLVFDARSSAGNMVIHGGPKSGKSTALQTFILSAASLHSPHEVSFYCLDYGGGQLRALQDLAHVGSVASALEPERIRRTFGELEQLLLSRQQREVFRDRGANGSTPDDGFGEVFLVIDNLYGFGRDNTDQFNTRNPLLARVTELVNVGLAYGIHVIITTPSWLEVPLAMRDGLGLRLELRLHDARDSNVRVVGALRRPADAVPHDQPGRGLTMAAEHFLFAAPELDAQTNPVAAINARYPGMAAPPVRLLPTNLAPHAVGELYRGPDQLVIGQREEDLAPVILDLAANPLLMVFGDARSGKTTLLRHIIRTVREHSTADRVAFTVLDRRLHLVDEPLFPDNEYTANIDRIIPAMLGLANLIEARRPPAGMSAAELSRWTFAGHTHYLIIDDVDQVPDSPAMTGPYIGQRPWTPLIGLLAQAGDLGLRVIVTGRATGSAHLLMTSPLLRRFNDLQATTLMLAGNPADSGKIRGERFARLPAGRAILLTDSDSPTYVQLINPLVDAAAVSGETQQKGSQS