Structural Insights of Transcriptionally Active, Full-Length Androgen Receptor Coactivator Complexes

May combine two claims.

AR mutants lacking AF-2 or splicing variants lacking the LBD still have potent transcription activation.

“AF-2 deletion mutants of AR or AR splicing variants lacking LBD still possess potent transcription activation functions (Guo et al., 2009; Jenster et al., 1995; Jenster et al., 1991; Simental et al., 1991).”

• cites Guo et al., 2009 sourceSupported
• cites Jenster et al., 1995 sourceNot checked — no full text of the source
• cites Jenster et al., 1991 sourcePartly supported

  In “Domains Required for Transcriptional Activation” (page unavailable), the paper reports that an AR mutant lacking the complete steroid-binding domain activated transcription at approximately the level of hormone-bound wild-type AR. It studied engineered deletion mutants, not AR splice variants, and does not specifically identify an AF-2 deletion. Those distinctions are not supported by this source. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

Classified as probably not a substantial claim.

The AR LBD and DBD X-ray crystal structures have been solved separately.

“The LBD and DBD X-ray crystal structures of AR have been solved separately (Matias et al., 2000; Sack et al., 2001; Shaffer et al., 2004).”

• cites Matias et al., 2000 sourceNot checked — no full text of the source
• cites Sack et al., 2001 sourceSupported
• cites Shaffer et al., 2004 sourceSupported

May combine two claims.

AR AF-1 is responsible for p300 recruitment because the majority of the AR-Ab1 binding region is AF-1 (amino acids 142-485).

“This suggests that AR AF-1 is responsible for p300 recruitment since majority of AR-Ab1 binding region is the AF-1 (aa 142–485) (McEwan, 2004).”

• cites McEwan, 2004 sourcePartly supported

  The Transcriptional activation section defines AR AF-1 as amino acids 142–485. The Protein-protein interactions section reports that AR acetylation enhances p300 binding, but it does not identify an AR-Ab1 binding region or establish that AF-1 is responsible for p300 recruitment. Page numbers are unavailable in our copy. (LLM, conf=0.83)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

An androgen-induced inter-molecular N/C interaction was observed, supporting an anti-parallel dimerization model.

“On the other hand, an androgen-induced inter-molecular N/C interaction was observed, supporting an anti-parallel dimerization model (Langley et al., 1998; Langley et al., 1995; van Royen et al., 2012).”

• cites Langley et al., 1998 sourceNot checked — no full text of the source
• cites Langley et al., 1995 sourceSupported
• cites van Royen et al., 2012 sourceNot checked — no full text of the source

May combine two claims.

The LBD-DBD domain arrangement is similar to the HNF4α dimer and different from the non-symmetric PPARγ-RXRα dimer or the X-shape LBD-DBD crossing arrangement in the RXRα-LXRβ dimer.

“This LBD-DBD domain arrangement is similar to the HNF4α dimer that was solved by X-ray crystallography (Chandra et al., 2013), which is different from the non-symmetric PPARγ-RXRα dimer (Chandra et al., 2008) or the X-shape LBD-DBD crossing arrangement in RXRα-LXRβ dimer (Lou et al., 2014).”

• cites Chandra et al., 2013 sourceSupported
• cites Chandra et al., 2008 sourceSupported
• cites Lou et al., 2014 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

The AR DBD forms a head-to-head dimer on DNA.

“On one hand, the AR DBD forms a head-to-head dimer on DNA (Shaffer et al., 2004).”

• cites Shaffer et al., 2004 sourceSupported

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

A symmetric dimeric AR LBD structure was solved recently, suggesting that AR may form a dimer similar to the way ERα dimerizes.

“A symmetric dimeric AR LBD structure was also solved recently (Nadal et al., 2017), suggesting that AR may form a dimer similar to the way ERα dimerizes.”

• cites Nadal et al., 2017 sourcePartly supported

  The Results section reports a symmetrical AR-LBD core dimer, supporting the structural finding. But the Discussion distinguishes its H5-mediated interface from the canonical ER-subclass dimerization mode, so the proposed similarity in how they dimerize is not supported. Page numbers are unavailable in our copy. (LLM, conf=0.87)

  Experimental — accuracy not yet measured

May combine two claims.

The AR NTD contains intrinsically disordered regions, and the isolated NTD exists as a collapsed disordered or molten globule-like structure that is intermediate between a folded globule and an unfolded random coil.

“Biochemical and computational analyses suggest that the AR NTD contains intrinsically disordered regions and the isolated NTD exists as a “collapsed” disordered or molten globule-like structure, which represents an intermediate state between a folded globule structure and an unfolded random-coil structure (Lavery and McEwan, 2008b; Reid et al., 2002).”

• cites Lavery and McEwan, 2008bNot matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

• cites Reid et al., 2002 sourcePartly supported

  The Discussion (page not provided) describes the studied AR transactivation region as partially unfolded, with regions of secondary structure; the Results also find limited secondary and tertiary structure in the isolated fragment. The paper does not characterize the whole isolated NTD as a collapsed disordered or molten globule-like structure, or explicitly place it between a folded globule and a random coil. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

Classified as probably not a substantial claim. May combine two claims.

P300 inhibitor directly inhibits AR transcriptional activity and H3K27ac deposition at ARE site.

“P300 inhibitor directly inhibits AR transcriptional activity and H3K27ac deposition at the ARE site (Lasko et al., 2017).”

• cites Lasko et al., 2017 sourcePartly supported

  The paper reports that A-485 attenuated AR-dependent gene expression and reduced H3K27Ac at the androgen response element of the PSA enhancer. It describes direct inhibition of AR transcriptional activity as a suggestion from the timing of those effects, rather than an established finding; the claim omits that uncertainty. (LLM, conf=0.84)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

The AR NTD has very little homology with other NR NTDs.

“It also shares very little homology with other NR NTDs (Lavery and McEwan, 2005).”

• cites Lavery and McEwan, 2005 sourceSupported

May combine two claims.

Antiandrogen-resistant AR mutations such as F876L likely cause LBD structural change and stabilize active NTD conformation, converting antiandrogen into agonist.

“On the other hand, antiandrogen-resistant AR mutations, such as an F876L mutation (Balbas et al., 2013), likely cause a LBD structural change, and consequently stabilize the active NTD conformation to convert an antiandrogen (e.g. enzalutamide) into an agonist.”

• cites Balbas et al., 2013 sourceSupported

May combine two claims.

Coactivator binding partners stabilize the NTD structure and convert it into a protease-resistant conformation.

“Interaction of the NTD with its coactivator binding partners stabilizes the NTD structure and converts the NTD into a protease-resistant conformation (Kumar et al., 2004; Lavery and McEwan, 2008b; Reid et al., 2002).”

• cites Kumar et al., 2004 sourceNot checked — no full text of the source
• cites Lavery and McEwan, 2008bNot matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

• cites Reid et al., 2002 sourcePartly supported

  The Discussion reports that binding of RAP74, a subunit of the general transcription factor TFIIF, promotes folding of the AR transactivation domain. The protein-interactions section reports that RAP74 binding makes the domain less susceptible to proteolysis. The paper tests this specific binding partner, not coactivator binding partners generally; page numbers are unavailable in our copy. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

May combine two claims.

The NTDs surround the LBDs to make both intra- and inter-molecular N/C interactions.

“Consistent with previous reports (Langley et al., 1998; Langley et al., 1995; van Royen et al., 2012), the NTDs also surround the LBDs to make both intra- and inter-molecular N/C interactions.”

• cites Langley et al., 1998 sourceNot checked — no full text of the source
• cites Langley et al., 1995 sourcePartly supported

  The Discussion supports an intermolecular interaction between the androgen-bound steroid-binding domain (LBD) and the NH2-terminal region (NTD). It says an intramolecular interaction cannot be ruled out, but does not establish one; nor does it show that the NTDs surround the LBDs. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites van Royen et al., 2012 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Gain-of-function ERα LBD mutations are frequently found in metastatic endocrine-resistant breast cancers and recruit elevated SRC coactivators to EREs.

“The gain-of-function ERα LBD mutations are frequently found in metastatic endocrine-resistant breast cancers (Jeselsohn et al., 2014; Robinson et al., 2013; Spoerke et al., 2016; Toy et al., 2013; Zhang et al., 1997) and recruit elevated levels of the SRC coactivators to EREs (Gates et al., 2018).”

• cites Jeselsohn et al., 2014 sourceSupported
• cites Robinson et al., 2013 sourceSupported
• cites Spoerke et al., 2016 sourceSupported

May combine two claims.

The SRC-3 RID pulls down ERα, consistent with previous reports.

“Consistent with previous reports (Ding et al., 1998; Heery et al., 1997), SRC-3 RID was found to pull-down ERα (Figure 4C).”

• cites Ding et al., 1998 sourceNot checked — no full text of the source
• cites Heery et al., 1997 sourceNot checked — no full text of the source

May combine two claims.

p300 directly interacts with AR without bridging through SRC-3, unlike the interaction in ERα.

“Further, we find p300 directly interacts with AR, without bridging through SRC-3, which is completely unlike that for ERα (Yi et al., 2015).”

• cites Yi et al., 2015Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

May combine two claims.

Full-length and 1-669aa AR fragments, but not the 600-920aa fragment, interact with p300.

“We observed that full-length and 1–669aa fragments, but not the 600–920aa fragment, of AR interacts with p300 (Figure 4D), confirming that the AR NTD is important for interaction with p300.”

May combine two claims.

Both NTDs connect to each other and contribute to AR dimerization, in addition to the LBD and DBD forming a tight dimerization interface at the center.

“In addition to LBD and DBD, which form a tight dimerization interface at the center, both NTDs also connect to each other to contribute to AR dimerization.”

May combine two claims.

p300 largely contacts the NTDs of both AR monomers and a small area also touches both LBDs.

“p300 largely contacts the NTDs of both AR monomers (-a and -b in Figure 3C). A small area of p300 also touches both LBDs.”

May combine two claims.

The resolved LBDs are located in the center of the AR dimer and sit on top of the DNA-bound DBDs.

“Our resolved LBDs are located in the center of the AR dimer and they sit on top of the DNA-bound DBDs (Figure 1B and Supplemental Figure 2G, 2H).”

May combine two claims.

The AR-Ab2-labeled AR dimer docks into the AR/coactivator complex such that the antibody is located next to the SRC-3 binding site, indicating that the SRC-3-interacting region is close to the AR N-terminal FXXLF motif where N/C interaction occurs.

“We also docked the AR-Ab2-labeled AR dimer structure into the AR/coactivator complex (Figure 2B) and found that this antibody is located right next to SRC-3 binding site (Supplemental Figure 5D, 5E.), indicating that the region interacting with SRC-3 is close to AR N-terminal FXXLF motif, where N/C interaction occurs.”

May combine two claims.

The bHLH/PAS, S/T, and HAT domains of SRC-3 each pulled down AR in vitro.

“Instead, the bHLH/PAS, S/T and HAT domains were each able to pull-down AR in vitro.”

May combine two claims.

ERα does not directly recruit p300, whereas AR interacts with p300 mainly through two NTDs.

“Third, ERα does not directly recruit p300, while AR interacts with p300 mainly through two NTDs.”

May combine two claims.

The paper's structural and protein-protein interaction studies indicate p300 directly interacts with AR, unlike its indirect interaction with ERα.

“Our structural and protein-protein interaction studies indicate that p300 directly interacts with AR in contrast to its indirect interaction with ERα.”

May combine two claims.

ERα recruits two SRC-3 molecules, whereas AR recruits only one SRC-3 molecule.

“First, ERα recruits two SRC-3 molecules, while AR only recruits one SRC-3 molecule.”

May combine two claims.

SRC-3 interacts with ERα LBD mainly via LXXLL motifs, but with AR NTD mainly through N-terminal regions.

“Second, SRC-3 mainly interacts with the ERα LBD via its LXXLL motifs, while it interacts with AR NTD mainly through its N-terminal regions.”

May combine two claims.

Androgen-bound AR has a strong AF-1 function and a relatively weak AF-2 function, unlike estrogen-bound estrogen receptor.

“Unlike estrogen-bound estrogen receptor, the androgen-bound AR has a strong AF-1 function and a relatively weak AF-2 function.”

May combine two claims.

Ligand binding causes the steroid receptor LBD to change conformation and expose an AF-2 hydrophobic groove that binds the LXXLL motif of coactivators such as SRCs.

“Upon ligand binding, the steroid receptor LBD undergoes a conformational change and presents a hydrophobic groove (AF-2) at the surface to interact with a LXXLL motif present in coactivators such as steroid receptor coactivators (SRCs) (Heery et al., 1997).”

• cites Heery et al., 1997 sourceNot checked — no full text of the source

Classified as probably not a substantial claim. May combine two claims.

The mechanism by which AR AF-1 dominates activation and how the N/C interaction contributes to coactivator recruitment and AR transcriptional activity is unclear.

“The underlying mechanism as to why the AF-1 of AR is its dominant activation domain and how its N/C interaction contributes to coactivator recruitment and AR transcriptional activity remains unclear.”

Probably not a substantial claim: may not pass S3 (load-bearing).

SRCs recruit additional coactivators, including p300/CBP histone acetyltransferases, to promote histone modification or chromatin remodeling and stimulate transcriptional initiation.

“SRCs, in turn, recruit other coactivators that are capable of promoting histone modifications or chromatin remodeling, such as the p300/CBP histone acetyltransferases, to stimulate transcriptional initiation.”

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based). May combine two claims.

All steroid hormone receptors form homodimers and bind palindromic DNA response elements upon ligand binding.

“All steroid hormone receptors form homodimers and bind to palindromic DNA response elements upon ligand binding.”

May combine two claims.

A strong N-terminal AF-1 function has been documented for AR, while a ligand-dependent AF-2 in the C-terminus is the major contributor to ERα activity.

“A strong N-terminal AF-1 function has been documented for AR, while a ligand-dependent AF-2 in the C-terminus is the major contributor to ERα activity (Jenster et al., 1995; Jenster et al., 1992; Kumar et al., 1987; Tora et al., 1989).”

• cites Jenster et al., 1995 sourceNot checked — no full text of the source
• cites Jenster et al., 1992 sourceNot checked — no full text of the source
• cites Kumar et al., 1987 sourceNot checked — no full text of the source

May combine two claims.

The coactivator assembly mechanism is different from ERα-mediated coactivator recruitment and implies that AR-recruited SRC-3 and p300 may have different conformations from ERα-recruited coactivators.

“This coactivator assembly mechanism is different from ERα-mediated coactivator recruitment (Yi et al., 2015) and implies that AR-recruited SRC-3 and p300 may have different conformations from ERα-recruited coactivators.”

• cites Yi et al., 2015Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

May combine two claims.

AR and ERα belong to the same steroid hormone receptor family and have similar domain organizations but utilize activation domains differently to initiate transcription.

“Although AR and ERα belong to the same steroid hormone receptor family and have similar domain organizations, they utilize activation domains differently to initiate transcription.”

May combine two claims.

The 3-D structure of full-length AR and its complex with SRC-3 and p300 elucidate differences between AR and ERα coactivator complex assembly and highlight the structural mechanism and importance of the AR NTD in transcription activation.

“The 3-D structure of full-length AR, as well as its complex structure with the two key coactivators presented here, elucidate the underlying differences between AR and ERα coactivator complex assembly and highlight the structural mechanism and importance of the AR NTD in transcription activation.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

The full-length ERα dimer forms a symmetric tail-to-tail dimer in the ERα/SRC-3/p300 complex, consistent with the palindromic DNA binding sequence and symmetric X-ray crystal structures of the ERα LBD and DBD dimers.

“We previously found that the full-length ERα dimer forms a symmetric tail-to-tail dimer in the ERα/SRC-3/p300 complex (Yi et al., 2015), consistent with the nature of palindromic DNA binding sequence, symmetric X-ray crystal structures of the ERα LBD (Brzozowski et al., 1997) and DBD dimers (Schwabe et al., 1993).”

• cites Yi et al., 2015Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

• cites Brzozowski et al., 1997 sourceNot checked — no full text of the source
• cites Schwabe et al., 1993 sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

The starting map was generated from scratch, and the overall structure was validated by subtomogram averaging from a different region of the same grid.

“The starting map of the refinement was generated from scratch and the overall structure was validated using subtomogram averaging performed in a different region of the same grid (Supplemental Figure 1F).”

Classified as probably not a substantial claim.

The precise structural role of AR interdomain interactions in full-length receptor dimerization and coactivator recruitment for transcription initiation remains elusive.

“The precise structural role of AR interdomain interactions in the context of full-length receptor dimerization and coactivator recruitment for regulating transcription initiation remains elusive.”

May combine two claims.

The two NTDs within the AR dimer display different modes of N/C interactions and adopt different conformations to form unique interaction surfaces for recruiting SRC-3 and p300, respectively.

“The two NTDs within the AR dimer display different modes of N/C interactions, and consequently adopt different conformations to form unique interaction surfaces for recruiting SRC-3 and p300 coactivators, respectively (Supplemental Figure 4C).”

May combine two claims.

The NTD not recognized by Ab2 overlaps more with the LBDs and is less exposed, suggesting it is more engaged in inter- and intra-molecular N/C interactions than the other NTD.

“The NTD not recognized by Ab2 shows more overlap with other segmented domains (the two LBDs) and has less exposed areas in the density, suggesting that it may be more engaged in both inter- and intra-molecular N/C interactions compared to the other NTD.”

May combine two claims.

Upon androgen binding, AR dimerizes and undergoes both intra- and inter-molecular N/C interactions.

“Upon androgen binding, AR dimerizes and undergoes both intra- and inter-molecular N/C interactions.”

May combine two claims.

The NTD, but not the LBD, is important for AR interaction with SRC-3.

“In contrast, the 600–920aa mutant has lost this interaction, suggesting that the NTD, but not the LBD, is important for the AR interaction with SRC-3.”

May combine two claims.

The volume ratio between each component agrees with the ratio of their molecular weights, further validating the segmentation.

“The volume ratio between each component is in agreement with the ratio of their molecular weights, further validating the segmentation.”

Probably not a substantial claim: may not pass S3 (load-bearing).

The mode of AR dimer formation is controversial.

“The mode of AR dimer formation, however, is controversial.”

May combine two claims.

The two antibody-bound density maps are consistent with the ARE-DNA/AR/SRC-3/p300 density map with an obvious protruding density, indicating the locations of specific targeting components.

“The two antibody-bound density maps are consistent with the ARE-DNA/AR/SRC-3/p300 density map with an obvious protruding density, which indicates the location of specific targeting components in the complex.”

May combine two claims.

The structural data agree with prior reports that AR NTD has strong activation function and that many coactivators including SRCs and CBP/p300 can be recruited through AR NTD.

“Our structural data is consistent with previous reports that the AR NTD has a strong activation function and that many coactivators, including SRCs and CBP/p300, can be recruited through its AR NTD (Alen et al., 1999; Jenster et al., 1995; Jenster et al., 1992; Lavery and McEwan, 2008a; Quayle et al., 2007).”

• cites Alen et al., 1999 sourceNot checked — no full text of the source
• cites Jenster et al., 1995 sourceNot checked — no full text of the source
• cites Jenster et al., 1992 sourceNot checked — no full text of the source

May combine two claims.

In the full-length receptor, AR NTDs are less disordered and likely stabilized by interaction with LBDs.

“suggesting that the NTDs in the context of full-length receptor are less disordered and likely stabilized by its interaction with the LBDs.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Error bars show the standard error of the mean.

“Statistical test used in this study was the Student’s t test using Excel. The n number is three independent experiments with duplicates in each experiment. The error bar represents standard error of the mean (SEM). p<0.05 was considered statistically significant.”

Probably not a substantial claim: may not pass S3 (load-bearing).

In the ERα tail-to-tail dimer, one NTD is on one side of the LBD and the other NTD is on the opposite side.

“It also is different from the ERα tail-to-tail dimer, which has a NTD located at one side of the LBD distant from the other NTD located at the opposite side (Yi et al., 2015).”

• cites Yi et al., 2015Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

May combine two claims.

The structural studies highlight AR NTD importance in coactivator recruitment and provide mechanistic insights into differences between ERα and AR mutations in breast and prostate cancer.

“Our structural studies presented here highlight the importance of the AR NTD in AR coactivator recruitment and provide mechanistic insights for observed differences between ERα and AR mutations in breast and prostate cancer.”

May combine two claims.

Without androgen or with AR antagonist, AR NTDs may adopt different conformations and have lower binding affinities for coactivators in full-length receptor.

“Without androgen stimulation or with AR antagonist treatment, AR NTDs may adopt different conformations and therefore have lower binding affinities with coactivators in the context of full-length receptor.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

The study used Student's t test and considered p<0.05 significant.

“Statistical test used in this study was the Student’s t test using Excel. The n number is three independent experiments with duplicates in each experiment. The error bar represents standard error of the mean (SEM). p<0.05 was considered statistically significant.”

May combine two claims.

Preclinical studies show p300 inhibitor efficacy in patient-derived prostate tumor explants and castration-resistant prostate cancer growth.

“Preclinical studies of a p300 inhibitor also reveal efficacy in patient-derived prostate tumor explants (Butler et al., 2019) and the growth of castration-resistant prostate cancer (Jin et al., 2017).”

• cites Butler et al., 2019 sourceNot checked — no full text of the source
• cites Jin et al., 2017 sourceNot checked — no full text of the source

May combine two claims.

NTD-a appears to adopt a more extended conformation and apparently mediates both intra- and inter-molecular N/C interactions.

“The NTD-a, which appears to adopt a more extended conformation and apparently mediates both intra- and inter-molecular N/C interactions, might not to be optimal for interaction with SRC-3.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Component ratios were checked against molecular weight ratios from earlier ER structures.

“The ratio between each component was validated by the molecular weight ratio as described in the previous ER structures (Yi et al., 2017; Yi et al., 2015).”

• cites Yi et al., 2017Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

• cites Yi et al., 2015Not matched

  Couldn’t be matched to a paper in Crossref, so it can’t be checked.

May combine two claims.

The relatively large AR NTDs wrap around LBDs and block SRC-3 access to LBD.

“The relatively large-sized AR NTDs, however, wrap around the LBDs, blocking the access of SRC-3 to the LBD.”

May combine two claims.

Through this interaction, the AR NTD covers most LBD surfaces and limits LBD accessibility to coactivators.

“Through this interaction the AR NTD covers most of LBD surfaces to limit the accessibility of LBD to coactivators.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Three independent experiments, each with duplicates, provided the sample size.

“Statistical test used in this study was the Student’s t test using Excel. The n number is three independent experiments with duplicates in each experiment. The error bar represents standard error of the mean (SEM). p<0.05 was considered statistically significant.”

May combine two claims.

Consequently, SRC-3 is recruited solely by AR NTD, and p300 recruitment is stabilized by contact with two AR NTDs.

“Consequently, SRC-3 is solely recruited by the AR NTD, and p300 recruitment is stabilized by contacting two AR NTDs.”

Probably not a substantial claim: may not pass S3 (load-bearing).

The AR NTD is less conserved across vertebrate species than other AR domains.

“The AR NTD is conserved less across different vertebrate species compared to other domains (van de Wijngaart et al., 2012).”

• cites van de Wijngaart et al., 2012 sourceNot checked — no full text of the source