Coaggregation of polyglutamine (polyQ) proteins is mediated by polyQ-tract interactions and impairs cellular proteostasis.

Nine polyglutamine (polyQ) proteins have already been identified that are considered to be associated with the pathologies of neurodegenerative disorders called polyQ diseases, but whether these polyQ proteins mutually interact and synergize in proteinopathies remains to be elucidated. In this study, 4 polyQ-containing proteins, androgen receptor (AR), ataxin-7 (Atx7), huntingtin (Htt) and ataxin-3 (Atx3), are used as model molecules to investigate their heterologous coaggregation and consequent impact on cellular proteostasis. Our data indicate that the N-terminal fragment of polyQ-expanded (PQE) Atx7 or Htt can coaggregate with and sequester AR and Atx3 into insoluble aggregates or inclusions through their respective polyQ tracts. In vitro coprecipitation and NMR titration experiments suggest that this specific coaggregation depends on polyQ lengths and is probably mediated by polyQ-tract interactions. Luciferase reporter assay shows that these coaggregation and sequestration effects can deplete the cellular availability of AR and consequently impair its transactivation function. This study provides valid evidence supporting the viewpoint that coaggregation of polyQ proteins is mediated by polyQ-tract interactions and benefits our understanding of the molecular mechanism underlying the accumulation of different polyQ proteins in inclusions and their copathological causes of polyQ diseases.

PolyQ-expanded proteins impair cellular proteostasis of ataxin-3 through sequestering the co-chaperone HSJ1 into aggregates.

Polyglutamine (polyQ) expansion of proteins can trigger protein misfolding and amyloid-like aggregation, which thus lead to severe cytotoxicities and even the respective neurodegenerative diseases. However, why polyQ aggregation is toxic to cells is not fully elucidated. Here, we took the fragments of polyQ-expanded (PQE) ataxin-7 (Atx7) and huntingtin (Htt) as models to investigate the effect of polyQ aggregates on the cellular proteostasis of endogenous ataxin-3 (Atx3), a protein that frequently appears in diverse inclusion bodies. We found that PQE Atx7 and Htt impair the cellular proteostasis of Atx3 by reducing its soluble as well as total Atx3 level but enhancing formation of the aggregates. Expression of these polyQ proteins promotes proteasomal degradation of endogenous Atx3 and accumulation of its aggregated form. Then we verified that the co-chaperone HSJ1 is an essential factor that orchestrates the balance of cellular proteostasis of Atx3; and further discovered that the polyQ proteins can sequester HSJ1 into aggregates or inclusions in a UIM domain-dependent manner. Thereby, the impairment of Atx3 proteostasis may be attributed to the sequestration and functional loss of cellular HSJ1. This study deciphers a potential mechanism underlying how PQE protein triggers proteinopathies, and also provides additional evidence in supporting the hijacking hypothesis that sequestration of cellular interacting partners by protein aggregates leads to cytotoxicity or neurodegeneration.

Domain interactions reveal auto-inhibition of the deubiquitinating enzyme USP19 and its activation by HSP90 in the modulation of huntingtin aggregation.

Ubiquitin-specific protease 19 (USP19) is a member of the deubiquitinating (DUB) enzymes that catalyze removing the ubiquitin signals from target proteins. Our previous research has demonstrated that USP19 up-regulates the protein level and aggregation of polyQ-expanded huntingtin through the involvement of heat shock protein 90 (HSP90). Here, we present solution structures of the CS1, CS2 and UbL domains of USP19 and structural insights into their domain interactions. We found that the tandem CS domains fold back to interact with the C-terminal USP domain (USPD) intra-molecularly that leads to inhibition of the catalytic core of USP19, especially CS1 interacts with the embedded UbL domain and CS2 does with the CH2 catalytic core. Moreover, CS2 specifically interacts with the NBD domain of HSP90, which can activate the DUB enzyme. A mechanism of auto-inhibition of USP19 and activation by HSP90 is proposed, on which USP19 modulates the protein level of polyQ-expanded huntingtin in cells. This study provides structural and mechanistic insights into the modulation of protein level and aggregation by USP19 with the assistance of HSP90.

Author Correction: Structural and dynamic studies reveal that the Ala-rich region of ataxin-7 initiates α-helix formation of the polyQ tract but suppresses its aggregation.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Structural and dynamic studies reveal that the Ala-rich region of ataxin-7 initiates α-helix formation of the polyQ tract but suppresses its aggregation.

Ataxin-7 (Atx7) is a disease-related protein associated with the pathogenesis of spinocerebellar ataxia 7, while its polyglutamine (polyQ) tract in N-terminus is the causative source of aggregation and proteinopathy. We investigated the structure, dynamics and aggregation properties of the N-terminal 62-residue fragment of Atx7 (Atx7-N) by biochemical and biophysical approaches. The results showed that the normal Atx7-N with a tract of 10 glutamines (10Q) overall adopts a flexible and disordered structure, but it may contain a short or small population of helical structure in solution. PolyQ expansion increases the α-helical propensity of the polyQ tract and consequently enhances its transformation into ß-sheet structures during amyloid aggregation. An alanine-rich region (ARR) just ahead of the polyQ tract forms a local and relatively stable α-helix. The ARR α-helix can initiate and stabilize helical formation of the following polyQ tract, but it may suppress aggregation of the polyQ-expanded Atx7-N both in vitro and in cell. Thus, the preceding ARR segment in Atx7-N may influence the dynamic structure and aggregation property of the polyQ tract and even determine the threshold of the pathogenic polyQ lengths. This study may gain structural and dynamic insights into amyloid aggregation of Atx7 and help us further understand the Atx7 proteinopathy based on polyQ expansion.

PolyQ-expanded huntingtin and ataxin-3 sequester ubiquitin adaptors hHR23B and UBQLN2 into aggregates via conjugated ubiquitin.

The components of ubiquitin (Ub)-proteasome system, such as Ub, Ub adaptors, or proteasome subunits, are commonly accumulated with the aggregated proteins in inclusions, but how protein aggregates sequester Ub-related proteins remains elusive. Using N-terminal huntingtin (Htt-N552) and ataxin (Atx)-3 as model proteins, we investigated the molecular mechanism underlying sequestration of Ub adaptors by polyQ-expanded proteins. We found that polyQ-expanded Htt-N552 and Atx-3 sequester endogenous Ub adaptors, human RAD23 homolog B (hHR23B) and ubiquilin (UBQLN)-2, into inclusions. This sequestration effect is dependent on the UBA domains of Ub adaptors and the conjugated Ub of the aggregated proteins. Moreover, polyQ-expanded Htt-N552 and Atx-3 reduce the protein level of xeroderma pigmentosum group C (XPC) by sequestration of hHR23B, suggesting that this process may cut down the available quantity of hHR23B and thus affect its normal function in stabilizing XPC. Our findings demonstrate that polyQ-expanded proteins sequester Ub adaptors or other Ub-related proteins into aggregates or inclusions through ubiquitination of the pathogenic proteins. This study may also provide a common mechanism for the formation of Ub-positive inclusions in cells.-Yang, H., Yue, H.-W., He, W.-T., Hong, J.-Y., Jiang, L.-L., Hu, H.-Y. PolyQ-expanded huntingtin and ataxin-3 sequester ubiquitin adaptors hHR23B and UBQLN2 into aggregates via conjugated ubiquitin.

HSP90 recognizes the N-terminus of huntingtin involved in regulation of huntingtin aggregation by USP19.

Huntington's disease (HD) is caused by aberrant expansion of polyglutamine (polyQ) in the N-terminus of huntingtin (Htt). Our previous study has demonstrated that HSP90 is involved in the triage decision of Htt, but how HSP90 recognizes and regulates Htt remains elusive. We investigated the interaction between HSP90 and the N-terminal fragments of Htt (Htt-N), such as the N-terminal 90-residue fragment (Htt-N90). Our results showed that HSP90 binds to the N-terminal extreme of Htt-N in a sequence just ahead of the polyQ tract. Structural integration of the middle and C-terminal domains of HSP90 is essential for interacting with Htt-N90, and the dimerization mediated by the C-terminal domain facilitates this interaction. Moreover, ubiquitin-specific protease 19 (USP19), a deubiquitinating enzyme interacting with HSP90, up-regulates the protein level of Htt-N90 and consequently promotes its aggregation, whereas disruption of the interaction between Htt-N90 and HSP90 attenuates the effect of USP19 on Htt-N90. Thus, HSP90 interacts with Htt-N90 on the N-terminal amphipathic α-helix, and then recruits USP19 to modulate the protein level and aggregation of Htt-N90. This study provides mechanistic insights into the recognition between HSP90 and the N-terminus of Htt, and the triage decision for the Htt protein by the HSP90 chaperone system.

The N-terminal dimerization is required for TDP-43 splicing activity.

TDP-43 is a nuclear factor that functions in promoting pre-mRNA splicing. Deletion of the N-terminal domain (NTD) and nuclear localization signal (NLS) (i.e., TDP-35) results in mislocalization to cytoplasm and formation of inclusions. However, how the NTD functions in TDP-43 activity and proteinopathy remains largely unknown. Here, we studied the structure and function of the NTD in inclusion formation and pre-mRNA splicing of TDP-43 by using biochemical and biophysical approaches. We found that TDP-43 NTD forms a homodimer in solution in a concentration-dependent manner, and formation of intermolecular disulfide results in further tetramerization. Based on the NMR structure of TDP-43 NTD, the dimerization interface centered on Leu71 and Val72 around the ß7-strand was defined by mutagenesis and size-exclusion chromatography. Cell experiments revealed that the N-terminal dimerization plays roles in protecting TDP-43 against formation of cytoplasmic inclusions and enhancing pre-mRNA splicing activity of TDP-43 in nucleus. This study may provide mechanistic insights into the physiological function of TDP-43 and its related proteinopathies.

Study of Protein Amyloid-Like Aggregates by Solid-State Circular Dichroism Spectroscopy.

Protein aggregation and amyloidogenesis are closely associated with the pathogenesis of neurodegenerative diseases. Elucidating the morphology and structure of the amyloid aggregates or fibrils is important for understanding the molecular mechanisms of these proteinopathies. This review article describes the general principle and establishment of solid-state circular dichroism (ssCD) spectroscopy, and discusses its application for the analysis of secondary structures of proteins or peptides in amyloids and structural transformation of these proteins or peptides during their amyloidogenic aggregation.

A study on HLA-DQB1 allele associated with genetic susceptibility to duodenal ulcer in Guangdong Hans / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the genetic susceptibility of HLA-DQB1 alleles to duodenal ulcer in Chinese Hans from Guangdong area around.</p><p><b>METHODS</b>Hundred and five patients with duodenal ulcer and hundred and five healthy controls were examined for HLA-DQB1 genotypes. HLA-DQB1 allele typing was carried out by polymerase chain reaction with sequence specific primers (PCR-SSP).</p><p><b>RESULTS</b>The allele frequency of HLA-DQB1*0602 in patients with duodenal ulcer (64.8%) was significantly higher than that in healthy controls (14.3%).</p><p><b>CONCLUSION</b>These findings suggest that HLA-DQB1*0602 is a susceptible gene to duodenal ulcer in Guangdong Hans of China. And at HLA-DQB1 site, there are immunogenetic differences between duodenal ulcer patients and healthy controls.</p>

Clinical, enteroscopic, and pathological characteristics of 796 cases of colorectal polyps / 中南大学学报(医学版)

OBJECTIVE@#To study the age, clinical, enteroscopic and pathological characteristics of colorectal polyps and factors affected polyp-carcinoma.@*METHODS@#We analyzed the clinical, enteroscopic and pathological characteristics of 7276 cases of colorectal polyps.@*RESULTS@#The incidence of colorectal polyps was 10.94%, including 521 men and 275 women. The rate of colorectal polyp was 82.29% in 30-69 year olds. The adenomatous, inflammatory, hyperplastic and juvenile polyps were 43.84%, 42.09%, 11.06% and 1.51%, respectively. Polypoid lesions were located at cecum 3.29%, ascending 11.88%, transverse 4.89%, descending 11.58%, sigmoid 26.05%, and rectum 42.32%. Thirty-five cases (4.4%) were found to have polpous canceration. The canceration rates in villous, mixed and tubular adenomas were 29.73%, 11.11%, and 4.86%. The rate of canceration seemed to depend on its dimensions, being 1.3%, 7.4%, and 25.6% for the 0.6 - 1.0 cm, 1.1 - 1.9 cm, and > or = 2.0 cm in size, respectively. Conclusion The ages between 30-69 tend to suffer from colorectal polyps. The incidence in the male is higher than that in the female. Colorectal polyps are more likely to locate in left colon. The common pathological types were adenomatous and inflammatory polyps. There is a high canceration of polyps in the left colon, villous adenomas and > or = 2.0 cm polyps. The broader the pedicles and the larger the diameters of polyps are, the higher the canceration rate. All of the colon polyps should be excised and undergo the pathological examination. Enteroscopic polypectomy helps prevent colorectal polpous canceration.

Features and clinic values of normal lumbar nerve root anatomy with CT on multiple plane reconstruction techniques at the same slice / 中华放射学杂志

Objective To explore features and clinic values of LNR anatomy with multiple planar reconstruction techniques with 16-slice spiral CT at the same slice.Methods The lumbar vertebrae with normal adults of 55 cases and 23 cases with abnormal ENR caused by 8 cases with protrusion of lumbar disc, 5 cases with spinal stenosis,4 cases with malignant tumor,5 cases with trauma and 1 case with lumbar TB confirmed by operation were scanned with 16-slice spiral CT made in American GE company in routine posture of the lumbar vertebrae,reconstructed LNR with UNIX system in workstation (ADW 4.1),and analyzed their normal and abnormal anatomic manifestations at the same slice.Results All of LNR can symmetrically showed on oblique and coronal planes according to different segments:one segment from L1 to L5(55,100% ),two segments: from L1 to L2,L2 to L3 and L3 to L4(55,100% ),three segments: from L1 to L3 (49,88%),from L2to L4(46,84% )and from L3 to L5(20,36% ),four segments: from L1 to L4 (15,27% )and five segments:(8,15% ),respectively.Each LNR,including their whole shapes of passage from starting to end,direction,size,shape,tension and peripheral relationship and so on can showed clearly on oblique and coronal planes and on other planes. However,the later planes can increase LNR but decreasing numbers of LNR and especially increase very long one LNR reconstruction.Primary manifestation of all diseases can be showed on oppressing along its walking line,meanwhile,20 cases with adhesion, 14 cases with displacement,13 cases atrophy and 9 cases with increasing diameter.Conclusions Image anatomy features of full LNR with 16-slice spiral CT with the multiple plane reconstruction techniques is very ideal ways at the same slice.It is a very valuable way to make diagnosis and treatment of LNR diseases.The concept of"road sing"and showing"at the same slice"of LNR are tried to rise from in order to make foundation for studying their image.