TBXT (Brachyury, T) is the founding member of the T-box family of sequence-specific transcription factors. Its N-terminal T-box domain binds a palindromic T-site DNA motif in cis-regulatory regions, and the protein acts in the nucleus mainly as a transcriptional activator of RNA polymerase II target genes, such as MESP1 in early cardiovascular progenitors. It can also bind other developmental transcription factors such as the homeodomain protein MIXL1 and dampen their activity. TBXT is expressed in the primitive streak, nascent mesoderm, tailbud and notochord and is required for posterior mesoderm formation, axial elongation and notochord development; loss-of-function variants cause sacral agenesis with vertebral anomalies, and duplication or overexpression is associated with chordoma and other tumours. Brachyury is the oldest T-box class: orthologs exist in the unicellular filasterean Capsaspora owczarzaki, in ichthyosporeans and in early-branching fungi (they have been lost from choanoflagellates), and the Capsaspora protein binds a near-identical DNA motif, whereas the restricted, cofactor-dependent target specificity of Brachyury arose with animals.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0000122 negative regulation of transcription by RNA polymerase II | ISS GO_REF:0000024 | KEEP AS NON CORE | Summary: Transferred from mouse T IDA (PMID:22164283): Brachyury represses Mixl1-dependent activation of the Gsc promoter. Reason: Supported by reporter assays in which Brachyury inhibits Mixl1 transactivation, but this is indirect (no T site in the Gsc promoter) and secondary to TBXT's activator role. Supporting Evidence: PMID:22164283 luciferase reporter experiments indicated that association of Mixl1 with Brachyury and related T-box factors inhibited the transactivating potential of Mixl1 on the Gsc and PdgfrΞ± promoters. |
| GO:0000785 chromatin | IBA GO_REF:0000033 | ACCEPT | Summary: IBA for chromatin from the T-box family node. Reason: A sequence-specific TF that binds cis-regulatory elements in vivo (ChIP) acts on chromatin. Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. |
| GO:0000785 chromatin | IDA PMID:21632880 Induction of MesP1 by Brachyury(T) generates the common mult... | ACCEPT | Summary: ChIP shows Brachyury occupancy of the MesP1 promoter. Reason: ChIP is direct evidence of chromatin association. Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. |
| GO:0000785 chromatin | ISA GO_REF:0000113 | ACCEPT | Summary: TFClass-based assignment of chromatin location for a DNA-binding TF. Reason: Consistent with ChIP evidence. Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. |
| GO:0000978 RNA polymerase II cis-regulatory region sequence-specific DNA binding | IBA GO_REF:0000033 | ACCEPT | Summary: IBA from the T-box family node PTN004251338: sequence-specific binding of RNA polymerase II cis-regulatory sites. Human TBXT has its own experimental support (it appears in WITH/FROM), and the T-box DNA-binding domain (residues 51-219) is conserved throughout the family. Reason: Binding of T-site elements in cis-regulatory regions is the defining biochemical activity of Brachyury and is ancestral to the T-box family; Capsaspora Brachyury has a near-identical motif. Node placement is sound. Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. PMID:24043797 Our results indicate that CoBra has a highly similar motif to that determined in the mouse Bra-homolog, called T ( Fig. 4 ) ( 28 , 34 β 36 ). |
| GO:0000978 RNA polymerase II cis-regulatory region sequence-specific DNA binding | IDA PMID:21632880 Induction of MesP1 by Brachyury(T) generates the common mult... | ACCEPT | Summary: EMSA, ChIP and reporter assays show Brachyury binds a T responsive element in the MesP1 proximal promoter. Reason: Direct experimental evidence of sequence-specific binding to a Pol II cis-regulatory region; the abstract states it directly. Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. |
| GO:0000978 RNA polymerase II cis-regulatory region sequence-specific DNA binding | IEA GO_REF:0000002 | ACCEPT | Summary: InterPro2GO mapping from the T-box domain (IPR001699). Reason: Consistent with experimental data and with the conserved T-box DNA-binding domain. Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. |
| GO:0000978 RNA polymerase II cis-regulatory region sequence-specific DNA binding | ISS GO_REF:0000024 | ACCEPT | Summary: Transferred from mouse T (P20293), whose source annotation is an IDA from the Mixl1 interaction study (PMID:22164283) and PMID:17584735. Reason: The activity is correct for human TBXT and independently supported by human/IDA and IBA rows. The Mixl1 study itself found no T sites in the Gsc promoter, so that particular source is weak, but the claim stands on other evidence. Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. PMID:22164283 Inspection of the Gsc promoter did not reveal consensus DNA binding sites for Brachyury [22], [23], suggesting that Brachyury was not directly binding to the Gsc promoter to regulate Mixl1 activity. |
| GO:0000981 DNA-binding transcription factor activity, RNA polymerase II-specific | IBA GO_REF:0000033 | ACCEPT | Summary: IBA from PTN004251338 (T-box family): RNA polymerase II-specific DNA-binding transcription factor activity. Reason: Core activity of TBXT, ancestral to the T-box family. GO:0001228 (activator) is more precise and is proposed as NEW below; this broader term remains correct. Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. PMID:22164283 Brachyury (T), the founding member of the T-box (Tbx) family, is a transcriptional activator and is expressed throughout the nascent mesoderm, tailbud and notochord |
| GO:0000981 DNA-binding transcription factor activity, RNA polymerase II-specific | IDA PMID:21632880 Induction of MesP1 by Brachyury(T) generates the common mult... | ACCEPT | Summary: Brachyury binds and activates the MesP1 promoter via a T element (reporter and ChIP). Reason: Direct evidence of sequence-specific Pol II transcription factor activity. Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. |
| GO:0000981 DNA-binding transcription factor activity, RNA polymerase II-specific | ISA GO_REF:0000113 | ACCEPT | Summary: Assigned from TFClass classification (6.5.1, T-box/Brachyury-related). Reason: Correct classification of a T-box DNA-binding transcription factor. Supporting Evidence: PMID:22164283 Brachyury (T), the founding member of the T-box (Tbx) family, is a transcriptional activator and is expressed throughout the nascent mesoderm, tailbud and notochord |
| GO:0001707 mesoderm formation | IBA GO_REF:0000033 | ACCEPT | Summary: IBA mesoderm formation from node PTN001681641 (Brachyury clade; donors mouse T and zebrafish ntla). Reason: Core developmental role: T is required for formation of posterior mesoderm, acting as the transcription factor that drives the mesodermal program. Node is within Metazoa, appropriate for an animal process. Supporting Evidence: PMID:8963900 T protein is vital for the formation of posterior mesoderm and axial development in all vertebrates. PMID:24253444 The T gene (brachyury gene) is the founding member of the T-box family of transcription factors and is vital for the formation and differentiation of the mesoderm and the axial development of all vertebrates. |
| GO:0001708 cell fate specification | IBA GO_REF:0000033 | KEEP AS NON CORE | Summary: IBA cell fate specification from a deep T-box family node (PTN000137774) with donors across T-box classes in fly, worm, fish and mouse. Reason: T-box factors specify mesodermal cell fates, so the term is reasonable, but it is broad; mesoderm formation and notochord development capture TBXT's role more precisely. Supporting Evidence: PMID:24253444 The T gene (brachyury gene) is the founding member of the T-box family of transcription factors and is vital for the formation and differentiation of the mesoderm and the axial development of all vertebrates. |
| GO:0001756 somitogenesis | IBA GO_REF:0000033 | KEEP AS NON CORE | Summary: IBA somitogenesis from the Brachyury clade (donors mouse T, zebrafish ntla). Reason: Mouse T mutants have abnormal posterior somites, consistent with a role in posterior paraxial mesoderm; a downstream consequence of TBXT's mesoderm role rather than a core function. Supporting Evidence: PMID:8963900 Brachyury mutant mice, which lack T protein, die in utero with abnormal notochord, posterior somites, and allantois. |
| GO:0003007 heart morphogenesis | IBA GO_REF:0000033 | KEEP AS NON CORE | Summary: IBA heart morphogenesis from PTN001681641 (donors include human TBXT IDA and zebrafish ntla). Reason: TBXT acts in early mesoderm upstream of cardiac progenitor specification (via MESP1); heart morphogenesis is an indirect, downstream outcome. Supporting Evidence: PMID:21632880 shRNA-based loss of Brachyury(T) causes a dramatic decrease in MesP1 expression accompanied by reduced cardiac markers in differentiating embryonic stem cells |
| GO:0003007 heart morphogenesis | IDA PMID:21632880 Induction of MesP1 by Brachyury(T) generates the common mult... | KEEP AS NON CORE | Summary: Brachyury induces MesP1 and is required for cardiac marker expression in differentiating ES cells. Reason: The data show TBXT upstream of cardiovascular progenitor specification, not participation in heart morphogenesis itself; kept as non-core rather than removed because the full text is not cached. Supporting Evidence: PMID:21632880 shRNA-based loss of Brachyury(T) causes a dramatic decrease in MesP1 expression accompanied by reduced cardiac markers in differentiating embryonic stem cells |
| GO:0003700 DNA-binding transcription factor activity | IEA GO_REF:0000002 | MODIFY | Summary: InterPro2GO mapping from T-box domain signatures to the generic DNA-binding transcription factor activity. Reason: Correct but less specific than the Pol II-specific term already supported experimentally; replace with GO:0000981. Proposed replacements: DNA-binding transcription factor activity, RNA polymerase II-specific Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. |
| GO:0003700 DNA-binding transcription factor activity | NAS PMID:8963900 The human homolog T of the mouse T(Brachyury) gene; gene str... | MODIFY | Summary: Author statement in the human T cloning paper that T encodes a transcription factor. Reason: Correct but generic; the Pol II-specific term GO:0000981 is better supported (IDA, IBA). Proposed replacements: DNA-binding transcription factor activity, RNA polymerase II-specific Supporting Evidence: PMID:8963900 We have cloned the human gene encoding the transcription factor T. |
| GO:0003714 transcription corepressor activity | ISS GO_REF:0000024 | KEEP AS NON CORE | Summary: Transferred from mouse T, whose IDA (PMID:22164283) is based on Brachyury binding the Mixl1 homeodomain and repressing Mixl1-driven Gsc reporter activity without binding the Gsc promoter. Reason: The Mixl1 study fits the corepressor definition (binding a DNA-binding TF and reducing its activity), and human Brachyury was shown to bind Mixl1 in the same work. The evidence is overexpression reporter assays and the activity is secondary to TBXT's role as a DNA-binding activator. Supporting Evidence: PMID:22164283 luciferase reporter experiments indicated that association of Mixl1 with Brachyury and related T-box factors inhibited the transactivating potential of Mixl1 on the Gsc and PdgfrΞ± promoters. PMID:22164283 Inspection of the Gsc promoter did not reveal consensus DNA binding sites for Brachyury [22], [23], suggesting that Brachyury was not directly binding to the Gsc promoter to regulate Mixl1 activity. |
| GO:0005634 nucleus | IBA GO_REF:0000033 | ACCEPT | Summary: IBA nucleus from the T-box family node PTN004251338. Reason: Nuclear localisation is expected and shown experimentally for human TBXT. Supporting Evidence: PMID:22611028 a single band at the expected molecular weight was observed in nuclear fractions derived from the primary tumor of two lung cancer patients evaluated |
| GO:0005634 nucleus | IDA PMID:22611028 Brachyury, a driver of the epithelial-mesenchymal transition... | ACCEPT | Summary: Brachyury detected in nuclear but not cytosolic fractions of human lung tumours, and by nuclear IHC staining. Reason: Direct evidence in human tissue. Supporting Evidence: PMID:22611028 a single band at the expected molecular weight was observed in nuclear fractions derived from the primary tumor of two lung cancer patients evaluated |
| GO:0005634 nucleus | IDA PMID:24253444 Mutations in the T (brachyury) gene cause a novel syndrome c... | ACCEPT | Summary: SAVA syndrome study characterising the H171R T-box variant; localisation data are in the full text (abstract only cached). Reason: Experimental annotation from a paper whose full text is not cached; consistent with all other evidence, so defer to the curator. Supporting Evidence: PMID:24253444 The homozygous mutation results in diminished DNA binding, increased cell growth, and interferes with the normal expression of genes involved in ossification, notochord maintenance and axial mesoderm development. |
| GO:0005634 nucleus | IEA GO_REF:0000120 | ACCEPT | Summary: Combined automated annotation (InterPro/UniProt subcellular location). Reason: Correct. Supporting Evidence: PMID:22611028 a single band at the expected molecular weight was observed in nuclear fractions derived from the primary tumor of two lung cancer patients evaluated |
| GO:0005654 nucleoplasm | IDA GO_REF:0000052 | ACCEPT | Summary: Human Protein Atlas immunofluorescence localisation to nucleoplasm. Reason: Consistent with nuclear TF function. Supporting Evidence: PMID:22611028 a single band at the expected molecular weight was observed in nuclear fractions derived from the primary tumor of two lung cancer patients evaluated |
| GO:0006355 regulation of DNA-templated transcription | IEA GO_REF:0000002 | MODIFY | Summary: InterPro2GO mapping to regulation of DNA-templated transcription. Reason: Correct but generic; the experimentally supported, more specific term is positive regulation of transcription by RNA polymerase II. Proposed replacements: positive regulation of transcription by RNA polymerase II Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. |
| GO:0006357 regulation of transcription by RNA polymerase II | IBA GO_REF:0000033 | ACCEPT | Summary: IBA regulation of transcription by RNA polymerase II. Reason: Correct; TBXT regulates Pol II target genes such as MESP1. Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. |
| GO:0007165 signal transduction | NAS PMID:8963900 The human homolog T of the mouse T(Brachyury) gene; gene str... | REMOVE | Summary: Author statement from the human T cloning paper. Reason: Nothing in the paper supports signal transduction; TBXT is a nuclear sequence-specific transcription factor, not a signal transducer. Non-experimental annotation without support. Supporting Evidence: PMID:8963900 We have cloned the human gene encoding the transcription factor T. |
| GO:0007498 mesoderm development | TAS PMID:8963900 The human homolog T of the mouse T(Brachyury) gene; gene str... | ACCEPT | Summary: Traceable author statement that T is vital for posterior mesoderm formation. Reason: Correct; mesoderm development is a core process for TBXT (parent of mesoderm formation). Supporting Evidence: PMID:8963900 T protein is vital for the formation of posterior mesoderm and axial development in all vertebrates. |
| GO:0008595 anterior/posterior axis specification, embryo | TAS PMID:8963900 The human homolog T of the mouse T(Brachyury) gene; gene str... | KEEP AS NON CORE | Summary: Author statement that T is vital for axial development in vertebrates. Reason: Brachyury is needed for posterior axis extension and axial mesoderm, but 'axis specification' (setting up the A/P polarity) overstates its role; kept as non-core. Supporting Evidence: PMID:8963900 T protein is vital for the formation of posterior mesoderm and axial development in all vertebrates. |
| GO:0045893 positive regulation of DNA-templated transcription | IEA GO_REF:0000002 | MODIFY | Summary: InterPro2GO (IPR046360) mapping to positive regulation of DNA-templated transcription. Reason: Correct direction; replace with the Pol II-specific child supported by IDA. Proposed replacements: positive regulation of transcription by RNA polymerase II Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. |
| GO:0045944 positive regulation of transcription by RNA polymerase II | IDA PMID:21632880 Induction of MesP1 by Brachyury(T) generates the common mult... | ACCEPT | Summary: Brachyury directly activates the MesP1 promoter in reporter assays. Reason: Direct evidence of activation of a Pol II promoter; Brachyury is primarily an activator. Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. PMID:22164283 Brachyury (T), the founding member of the T-box (Tbx) family, is a transcriptional activator and is expressed throughout the nascent mesoderm, tailbud and notochord |
| GO:0060379 cardiac muscle cell myoblast differentiation | IDA PMID:21632880 Induction of MesP1 by Brachyury(T) generates the common mult... | KEEP AS NON CORE | Summary: Knockdown of Brachyury reduces MesP1 and cardiac markers in ES cell differentiation. Reason: Supported upstream role in cardiovascular progenitor generation; not a core function of TBXT. Supporting Evidence: PMID:21632880 shRNA-based loss of Brachyury(T) causes a dramatic decrease in MesP1 expression accompanied by reduced cardiac markers in differentiating embryonic stem cells |
| GO:0061629 RNA polymerase II-specific DNA-binding transcription factor binding | IPI PMID:22164283 Brachyury and related Tbx proteins interact with the Mixl1 h... | KEEP AS NON CORE | Summary: Human Brachyury binds MIXL1 (Q9H2W2) via their DNA-binding domains in pull-down assays. Reason: Supported interaction with a partner homeodomain TF; informative about cofactor-dependent specificity, but not the core activity. Supporting Evidence: PMID:22164283 Here we report that Mixl1 interacts physically and functionally with the T-box protein Brachyury and related members of the T-box family of transcription factors. |
| GO:0061629 RNA polymerase II-specific DNA-binding transcription factor binding | ISS GO_REF:0000024 | KEEP AS NON CORE | Summary: Transferred from mouse T IPI with Mixl1/Gsc (PMID:22164283). Reason: Same evidence base as the human IPI row; redundant but correct. Supporting Evidence: PMID:22164283 Here we report that Mixl1 interacts physically and functionally with the T-box protein Brachyury and related members of the T-box family of transcription factors. |
| GO:0090009 primitive streak formation | NAS PMID:18593560 Mesp1 acts as a master regulator of multipotent cardiovascul... | KEEP AS NON CORE | Summary: Non-traceable author statement attributed to the Mesp1 master-regulator paper; the cached abstract does not mention Brachyury. Reason: Brachyury is expressed in the primitive streak and mouse mutants have posterior mesoderm defects, so the term is plausible. The cited source cannot be checked (abstract only), so it is kept as non-core rather than accepted. Supporting Evidence: PMID:22164283 Mixl1 and Brachyury are both expressed in the primitive streak PMID:18593560 During embryonic development, multipotent cardiovascular progenitor cells are specified from early mesoderm. |
| GO:1990837 sequence-specific double-stranded DNA binding | IDA PMID:28473536 Impact of cytosine methylation on DNA binding specificities ... | ACCEPT | Summary: High-throughput methylation-sensitive SELEX of human TFs (Yin et al. 2017); TBXT motif data are in the supplementary tables, not the cached main text. Reason: In vitro SELEX directly measures sequence-specific double-stranded DNA binding. Deferring to the curator who read the supplementary data; consistent with all other evidence. Supporting Evidence: PMID:28473536 By analysis of 542 human TFs with methylation-sensitive SELEX (systematic evolution of ligands by exponential enrichment), we found that there are also many TFs that prefer CpG-methylated sequences. |
| GO:1990837 sequence-specific double-stranded DNA binding | IEA GO_REF:0000117 | ACCEPT | Summary: ARBA machine-learning model annotation. Reason: Correct; the T-box domain binds a specific DNA motif. Less informative than GO:0000978, but not wrong. Supporting Evidence: PMID:24043797 Our results indicate that CoBra has a highly similar motif to that determined in the mouse Bra-homolog, called T ( Fig. 4 ) ( 28 , 34 β 36 ). |
| GO:0001228 DNA-binding transcription activator activity, RNA polymerase II-specific | IDA PMID:21632880 Induction of MesP1 by Brachyury(T) generates the common mult... | NEW | Summary: Brachyury binds a T responsive element in the MesP1 promoter and activates it in reporter assays; it is described as a transcriptional activator. Reason: More precise MF than GO:0000981 for TBXT, whose characterised direct targets are activated. Supporting Evidence: PMID:21632880 We finally defined a 3.4 kb proximal MesP1-promoter fragment which is directly bound and activated by Brachyury(T) via a T responsive element as shown via bandshift, chromatin immuneprecipitation, and reporter assays. PMID:22164283 Brachyury (T), the founding member of the T-box (Tbx) family, is a transcriptional activator and is expressed throughout the nascent mesoderm, tailbud and notochord |
| GO:0030903 notochord development | IMP PMID:24253444 Mutations in the T (brachyury) gene cause a novel syndrome c... | NEW | Summary: Human homozygous T-box variant H171R reduces DNA binding and causes a persistent notochordal canal; mouse T mutants have an abnormal notochord. Reason: TBXT is the transcription factor that drives the notochord gene program (participation as the regulator of the process, as for other developmental TFs). Comparator check: mouse T (P20293) carries GO:0030903 by IMP from three MGI papers, so the human absence is a transfer gap, not a convention. Supporting Evidence: PMID:24253444 The homozygous mutation results in diminished DNA binding, increased cell growth, and interferes with the normal expression of genes involved in ossification, notochord maintenance and axial mesoderm development. PMID:8963900 Brachyury mutant mice, which lack T protein, die in utero with abnormal notochord, posterior somites, and allantois. |
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Download this section (compressed HTML)Q: What is the endogenous function of Brachyury in the unicellular Capsaspora owczarzaki, which has no mesoderm or blastopore? Does it regulate a life-cycle transition (e.g. aggregative stage), and which genes does it bind in vivo?
Q: Which cofactors (e.g. Smad1) and which N- and C-terminal features confer the restricted target specificity of metazoan Brachyury that Capsaspora Brachyury lacks, and can they be mapped onto human TBXT?
Q: Should IBA process terms on the Brachyury clade (mesoderm formation, somitogenesis, heart morphogenesis) be placed at a Metazoa-or-later node, so that they do not propagate to Capsaspora or ichthyosporean Brachyury orthologs?
Q: Is the corepressor activity of Brachyury toward MIXL1 seen at endogenous levels in human primitive-streak-like cells, or only under overexpression?
Experiment: ChIP-seq or CUT&RUN of endogenous Capsaspora Brachyury across its life stages, compared with human TBXT ChIP-seq in mesoderm-differentiating hESCs, to test whether shared motif binding translates into overlapping targets.
Hypothesis: Capsaspora Brachyury binds T-sites broadly without the restricted, cofactor-dependent target set of human TBXT.
Experiment: Domain-swap chimeras of human TBXT and Capsaspora Brachyury expressed in TBXT-null hESC-derived mesoderm, with RNA-seq readout.
Hypothesis: Human N- and C-terminal regions, not the T-box, confer human-like target specificity.
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