Journal of Biological Chemistry最新文献_第9页

WWP1 mediates the ubiquitination and degradation of HIPK3 in bladder cancer cells. WWP1介导膀胱癌细胞中HIPK3的泛素化和降解。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-04-23 DOI: 10.1016/j.jbc.2025.108528

Haichao Chen,Xinyu Na,Pengcheng Hu,Qi Ma,Rui Yu

引用次数: 0

DNA stimulates the deacetylase SIRT6 to mono-ADP-ribosylate proteins with histidine repeats. DNA刺激去乙酰化酶SIRT6，使其与组氨酸重复序列形成单adp -核糖基化蛋白。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-04-23 DOI: 10.1016/j.jbc.2025.108532

Nicholas J Pederson, Katharine L Diehl

{"title":"DNA stimulates the deacetylase SIRT6 to mono-ADP-ribosylate proteins with histidine repeats.","authors":"Nicholas J Pederson, Katharine L Diehl","doi":"10.1016/j.jbc.2025.108532","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108532","url":null,"abstract":"Sirtuins are the NAD+-dependent class III lysine deacylases (KDACs). Members of this family have been linked to longevity and a wide array of different diseases, motivating the pursuit of sirtuin modulator compounds. Sirtuin 6 (SIRT6) is a primarily nuclear KDAC that deacetylates histones to facilitate gene repression. In addition to this canonical post-translational modification (PTM) \"eraser\" function, SIRT6 can use NAD+ instead to \"write\" mono-ADP-ribosylation (mARylation) on target proteins. This enzymatic function has been primarily associated with SIRT6's role in the DNA damage response. This modification has been challenging to study because it is not clear under what precise cellular contexts it occurs, only a few substrates are known, and potential interference from other ADP-ribosyltransferases in cells, among other reasons. In this work, we used commercially available ADP-ribosylation detection reagents to investigate the mARylation activity of SIRT6 in a reconstituted system. We observed that SIRT6 is activated in its mARylation activity by binding to dsDNA ends. We further identified a surprising target motif within biochemical substrates of SIRT6, polyhistidine (polyHis) repeat tracts, that are present in several previously identified SIRT6 mARylation substrates. This work provides important context for SIRT6 mARylation activity, in contrast to its KDAC activity, and generates a list of new potential SIRT6 mARylation substrates based on the polyHis motif..","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"108532"},"PeriodicalIF":4.0,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144001266","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

In situ detection of activation of CAPN3, a responsible gene product for LGMDR1, in mouse skeletal myotubes. 小鼠骨骼肌管中LGMDR1的负责基因产物CAPN3的原位检测。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-04-23 DOI: 10.1016/j.jbc.2025.108536

Chihiro Hisatsune,Fumiko Shinkai-Ouchi,Shoji Hata,Yasuko Ono

{"title":"In situ detection of activation of CAPN3, a responsible gene product for LGMDR1, in mouse skeletal myotubes.","authors":"Chihiro Hisatsune,Fumiko Shinkai-Ouchi,Shoji Hata,Yasuko Ono","doi":"10.1016/j.jbc.2025.108536","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108536","url":null,"abstract":"CAPN3/calpain-3/p94, a muscle-specific Ca2+-dependent cysteine protease, is responsible for limb-girdle muscular dystrophy R1 (LGMDR1), an autosomal recessive muscular dystrophy. However, the activation mechanism and physiological function of CAPN3 in skeletal muscles remain unknown. Here, we capture the in situ activation of CAPN3 in cultured mouse skeletal myotubes. Using our newly developed antibody, which specifically recognizes CAPN3 autolytic processing, we succeeded in differentiating wild-type CAPN3 from a protease-inactive CAPN3 mutant by immunostaining. We further demonstrated that CAPN3 predominantly localized at the M-bands of cultured skeletal myotubes at rest and translocated to the cytoplasm after activation by stimulation with ouabain, a cardiotonic steroid. This event requires a small but long-lasting cytoplasmic increase in Ca2+ levels, which is sufficient for the activation of CAPN3 but not of calpain-1/CAPN1. Activated CAPN3 digests the cytoskeletal proteins spectrin and talin. Thus, we successfully visualized the intracellular dynamics of endogenous CAPN3 in cultured skeletal muscles after activation by ouabain and demonstrated the subsequent processing of endogenous substrates in living cells. Our study will help understand the physiological functions of CAPN3 in skeletal muscles and the pathophysiological mechanisms of LGMDR1.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"81 1","pages":"108536"},"PeriodicalIF":4.8,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143885105","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Tamoxifen induces protection against manganese toxicity by REST upregulation via the ER-α/Wnt/β-catenin pathway in neuronal cells. 他莫昔芬通过神经元细胞ER-α/Wnt/β-catenin通路上调REST，诱导对锰毒性的保护。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-04-23 DOI: 10.1016/j.jbc.2025.108529

Alexis Digman,Edward Pajarillo,Sanghoon Kim,Itunu Ajayi,Deok-Soo Son,Michael Aschner,Eunsook Lee

{"title":"Tamoxifen induces protection against manganese toxicity by REST upregulation via the ER-α/Wnt/β-catenin pathway in neuronal cells.","authors":"Alexis Digman,Edward Pajarillo,Sanghoon Kim,Itunu Ajayi,Deok-Soo Son,Michael Aschner,Eunsook Lee","doi":"10.1016/j.jbc.2025.108529","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108529","url":null,"abstract":"Chronic exposure to elevated levels of manganese (Mn) causes a neurological disorder referred to as manganism, with symptoms resembling Parkinson's disease (PD). The repressor element-1 silencing transcription factor (REST) has been shown to be neuroprotective in several neurological disorders, including PD, suggesting that identifying REST upregulation mechanisms is an important avenue for the development of novel therapeutics. 17β-estradiol (E2) activates the Wnt/β-catenin signaling, which is known to increase REST transcription. E2 and tamoxifen (TX), a selective estrogen receptor modulator, exerted protection against Mn toxicity. In this study, we tested if TX upregulates REST potentially via Wnt/β-catenin signaling in Cath.a-differentiated (CAD) neuronal cells using luciferase assay, qPCR, western blot analysis, immunocytochemistry, mutagenesis, chromatin immunoprecipitation, and electrophoretic mobility shift assay. TX (1 μM) increased REST promoter activities and mRNA/protein levels and attenuated Mn-decreased REST transcription in parallel with TX's protective effects against Mn (250 μM)-induced toxicity, potentially via Wnt. TX activated Wnt/β-catenin signaling by preventing β-catenin degradation via inactivation of glycogen synthase kinase-3 beta, leading to increased β-catenin levels and its nuclear translocation and binding to T-cell factor/lymphoid enhancer binding factor sites on Wnt- responsive elements (WRE) of the REST promoter. Mutation of WRE abolished TX-induced REST promoter activity. TX-induced Wnt signaling activation was primarily via the estrogen receptor (ER)-α, although ER-β and G protein-coupled estrogen receptor 1 also mediated TX's action on REST transcription. These findings underscore the critical role of Wnt/β-catenin signaling in TX-induced REST transcription, affording protection mechanisms against Mn toxicity and neurological disorders associated with REST dysfunction.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"44 1","pages":"108529"},"PeriodicalIF":4.8,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143885108","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The mechanism of peptidoglycan O-acetylation in Gram-negative bacteria typifies bacterial MBOAT-SGNH acyltransferases. 革兰氏阴性菌中肽聚糖o -乙酰化的机制表征了细菌MBOAT-SGNH酰基转移酶的类型。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-04-23 DOI: 10.1016/j.jbc.2025.108531

Alexander C Anderson, Bailey J Schultz, Eric D Snow, Ashley S Brott, Stefen Stangherlin, Tyler Malloch, Jalen R London, Suzanne Walker, Anthony J Clarke

{"title":"The mechanism of peptidoglycan O-acetylation in Gram-negative bacteria typifies bacterial MBOAT-SGNH acyltransferases.","authors":"Alexander C Anderson, Bailey J Schultz, Eric D Snow, Ashley S Brott, Stefen Stangherlin, Tyler Malloch, Jalen R London, Suzanne Walker, Anthony J Clarke","doi":"10.1016/j.jbc.2025.108531","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108531","url":null,"abstract":"Bacterial cell envelope polymers are commonly modified with acyl groups that provide fitness advantages. Many polymer acylation pathways involve pairs of membrane-bound O-acyltransferase (MBOAT) and SGNH family proteins. As an example, the MBOAT protein PatA and the SGNH protein PatB are required in Gram-negative bacteria for peptidoglycan O-acetylation. The mechanism for how MBOAT-SGNH transferases move acyl groups from acyl-CoA donors made in the cytoplasm to extracellular polymers is unclear. Using the peptidoglycan O-acetyltransferase proteins PatAB, we explore the mechanism of MBOAT-SGNH pairs. We find that the MBOAT protein PatA catalyzes auto-acetylation of an invariant Tyr residue in its conserved C-terminal hexapeptide motif. We also show that PatB can use a synthetic hexapeptide containing an acetylated tyrosine to donate an acetyl group to a peptidoglycan mimetic. Finally, we report the structure of PatB, finding that it has structural features that shape its activity as an O-acetyltransferase and distinguish it from other SGNH esterases and hydrolases. Taken together, our results support a model for peptidoglycan acylation in which a tyrosine-containing peptide at the MBOAT's C-terminus shuttles an acyl group from the MBOAT active site to the SGNH active site, where it is transferred to peptidoglycan. This model likely applies to other systems containing MBOAT-SGNH pairs, such as those that O-acetylate alginate, cellulose, and secondary cell wall polysaccharides.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"108531"},"PeriodicalIF":4.0,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144001268","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Evidence of Bidirectional Transmembrane Signaling by the Sensor Histidine Kinase GacS from Pseudomonas aeruginosa. 铜绿假单胞菌组氨酸激酶传感器GacS双向跨膜信号传导的证据。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-04-23 DOI: 10.1016/j.jbc.2025.108521

Safoura Salar,Steve Silletti,Florian D Schubot

{"title":"Evidence of Bidirectional Transmembrane Signaling by the Sensor Histidine Kinase GacS from Pseudomonas aeruginosa.","authors":"Safoura Salar,Steve Silletti,Florian D Schubot","doi":"10.1016/j.jbc.2025.108521","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108521","url":null,"abstract":"Membrane-embedded signaling Histidine Kinases (SKs) from two-component and phosphorelay signal transduction systems play central roles in the gene regulation of bacteria, fungi, and plants. The SK GacS is a global regulator of gene expression in the human pathogen Pseudomonas aeruginosa. The interactions between GacS and another SK, RetS, are a model for studying non-canonical crosstalk in multikinase networks. During planktonic growth, RetS inhibits GacS to upregulate expression of virulence factors associated with acute P. aeruginosa infections and repress genes linked to chronic infection. Conversely, GacS activation promotes biofilm formation and chronic infection but suppresses factors required during acute infection. Using a combination of hydrogen-deuterium exchange mass spectrometry (HDX-MS) and mutational analysis in conjunction with functional assays, we show that binding of an extracellular ligand promotes GacS signaling through two mechanisms: (1) by increasing GacS autokinase activity and (2) by decreasing the affinity between GacS and RetS. Intriguingly, RetS binding to the intracellular histidine kinase domain of GacS also triggered conformational changes in the extracellular sensory domain of GacS. This allosteric effect was confirmed in a biochemical assay, showing RetS increases the affinity of a chimeric CitAGacS receptor for citrate by almost tenfold. This finding establishes the first precedent of inside-out cross-membrane signaling in SK systems. Taken together, our data are consistent with a model wherein RetS binding primes GacS for signal sensing during planktonic growth. Binding of the unknown ligand at the onset of biofilm formation causes dissociation of the RetS-GacS complex to lock GacS in a kinase ON conformation.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"274 1","pages":"108521"},"PeriodicalIF":4.8,"publicationDate":"2025-04-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143885109","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Aggrecan immobilizes to perineuronal nets through hyaluronan-dependent and independent binding activities. Aggrecan通过透明质酸依赖和独立的结合活动固定在神经周围网络上。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-04-22 DOI: 10.1016/j.jbc.2025.108525

Matthew Y Otsuka,Leslie B Essel,Ashis Sinha,Gabrielle Nickerson,Seth M Mejia,Ashley Edge,Russell T Matthews,Samuel Bouyain

{"title":"Aggrecan immobilizes to perineuronal nets through hyaluronan-dependent and independent binding activities.","authors":"Matthew Y Otsuka,Leslie B Essel,Ashis Sinha,Gabrielle Nickerson,Seth M Mejia,Ashley Edge,Russell T Matthews,Samuel Bouyain","doi":"10.1016/j.jbc.2025.108525","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108525","url":null,"abstract":"Aggrecan (ACAN) is a large, secreted chondroitin sulfate proteoglycan that includes three globular regions named G1, G2, G3, and is decorated with multiple glycosaminoglycan attachments between its G2 and G3 domains. The N-terminal G1 region interacts with the glycosaminoglycan hyaluronan (HA), which is an essential component of the vertebrate extracellular matrix. In the central nervous system, ACAN is found in perineuronal nets (PNNs), honeycomb-like structures that localize to the surface of parvalbumin-positive neurons in specific neural circuits. PNNs regulate the plasticity of the central nervous system, and it is believed that association between ACAN and HA is a foundational event in the assembly of these reticular structures. Here, we report the co-crystal structure of the G1 region of ACAN in the absence and presence of a HA decasaccharide and analyze the importance of the HA-binding activity of ACAN for its integration into PNNs. We demonstrate that the single immunoglobulin domain and the two Link modules that comprise the G1 region form a single structural unit, and that HA is clamped inside a groove that spans the length of the tandem Link domains. Introducing point mutations in the glycosaminoglycan-binding site eliminates HA-binding activity in ACAN, but, surprisingly, only decreases the integration of ACAN into PNNs. Thus, these results suggest that ACAN can be recruited into PNNs independently of its HA-binding activity.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"33 1","pages":"108525"},"PeriodicalIF":4.8,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143876478","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The ubiquitin ligase NKLAM promotes apoptosis and suppression of cell growth. 泛素连接酶NKLAM促进细胞凋亡和抑制细胞生长。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-04-22 DOI: 10.1016/j.jbc.2025.108527

Paul A Willard,Jacki Kornbluth

{"title":"The ubiquitin ligase NKLAM promotes apoptosis and suppression of cell growth.","authors":"Paul A Willard,Jacki Kornbluth","doi":"10.1016/j.jbc.2025.108527","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108527","url":null,"abstract":"Natural Killer Lytic Associated Molecule (NKLAM), also known as RNF19b, is a member of the RING-in between-RING-RING (RBR) E3 ubiquitin ligase family and plays a pivotal role in immune regulation. We identified a critical cysteine residue at position 301 essential for NKLAM's ubiquitin ligase function. Site-directed mutagenesis of this residue to serine or alanine abrogated the ligase activity of NKLAM. Utilizing inducible expression systems in two different cell lines, HEK293 embryonic kidney cells and K562 myeloid leukemia cells, we demonstrated that wild-type (WT) NKLAM, but not the catalytically inactive NKLAM alanine mutant (C301A), inhibited cellular proliferation, as evidenced by reduced cell numbers and decreased metabolic activity. Moreover, NKLAM expression led to a significant decrease in the abundance and stability of the proto-oncogene c-Myc, a key regulator of proliferation. NKLAM facilitated the proteasomal degradation of c-Myc, with a reduction in c-Myc half-life from 27 minutes to 12 minutes and restoration of c-Myc levels upon proteasome inhibition. Notably, prolonged NKLAM expression induced apoptosis, measured by annexin-V staining and caspase activation. Strikingly, the serine mutant, C301S, while lacking ubiquitin ligase activity, induced apoptosis comparable to WT NKLAM, highlighting an alternative pathway for NKLAM-mediated inhibition of cellular homeostasis. Our findings indicate that NKLAM is a cytolytic protein with multifaceted roles in cellular proliferation and apoptosis.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"76 1","pages":"108527"},"PeriodicalIF":4.8,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143876483","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Variations in kinase and effector signaling logic in a bacterial two component signaling network. 细菌双组分信号网络中激酶和效应信号逻辑的变化。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-04-22 DOI: 10.1016/j.jbc.2025.108534

Danielle Swingle, Leah Epstein, Ramisha Aymon, Eta A Isiorho, Rinat R Abzalimov, Denize C Favaro, Kevin H Gardner

{"title":"Variations in kinase and effector signaling logic in a bacterial two component signaling network.","authors":"Danielle Swingle, Leah Epstein, Ramisha Aymon, Eta A Isiorho, Rinat R Abzalimov, Denize C Favaro, Kevin H Gardner","doi":"10.1016/j.jbc.2025.108534","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108534","url":null,"abstract":"The general stress response (GSR) protects bacteria from a wide range of stressors. In Alphaproteobacteria, GSR activation is coordinated by HWE/HisKA2 family histidine kinases (HKs), which can exhibit non-canonical structure and function. For example, while most light-oxygen-voltage sensor-containing HKs are light activated dimers, the Rubellimicrobium thermophilum RT-HK has inverted \"dark on, light off\" signaling logic with a tunable monomer/dimer equilibrium. Here, we further investigate these atypical behaviors of RT-HK and characterize its downstream signaling network. Using hydrogen-deuterium exchange mass spectrometry, we find that RT-HK uses a signal transduction mechanism similar to light-activated systems, despite its inverted logic. Mutagenesis reveals that RT-HK autophosphorylates in trans, with changes to the Jα helix linking sensor and kinase domains affecting autophosphorylation levels. Exploring downstream effects of RT-HK, we identified two GSR genetic regions, each encoding a copy of the central regulator PhyR. In vitro measurements of phosphotransfer from RT-HK to the two putative PhyRs revealed that RT-HK signals only to one, and does so at an increased intensity in the dark, consistent with its reversed logic. X-ray crystal structures of both PhyRs revealed a substantial shift within the receiver domain of one, suggesting a basis for RT-HK specificity. We probed further down the pathway using nuclear magnetic resonance to determine that the single NepR homolog interacts with both unphosphorylated PhyRs, and this interaction is decoupled from activation in one PhyR. This work expands our understanding of HWE/HisKA2 family signal transduction, revealing marked variations from signaling mechanisms previously identified in other GSR networks.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"108534"},"PeriodicalIF":4.0,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144026167","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Phosphorylation modulates secondary structure of intrinsically disorder regions in RNA polymerase II. 磷酸化调节RNA聚合酶II内在紊乱区域的二级结构。

IF 4.8 2区生物学

Journal of Biological Chemistry Pub Date : 2025-04-22 DOI: 10.1016/j.jbc.2025.108533

Wei Chen,Tatiana N Laremore,Neela H Yennawar,Scott A Showalter

{"title":"Phosphorylation modulates secondary structure of intrinsically disorder regions in RNA polymerase II.","authors":"Wei Chen,Tatiana N Laremore,Neela H Yennawar,Scott A Showalter","doi":"10.1016/j.jbc.2025.108533","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108533","url":null,"abstract":"The intrinsically disordered C-terminal domain (CTD) of RNA polymerase II contains tandem repeats with the consensus sequence YSPTSPS and coordinates transcription and co-transcriptional events through dynamic phosphorylation patterns. While it has been long hypothesized that phosphorylation induces structural changes in the CTD, a direct comparison of how different phosphorylation patterns modulate the CTD conformation has been limited. Here, we generated two distinct phosphorylation patterns in an essential Drosophila CTD region with the kinase Dyrk1a: one where Ser2 residues are primarily phosphorylated, mimicking the state near transcription termination, and a hyperphosphorylation state where most Ser2, Ser5, and Thr residues are phosphorylated, expanding on our work on Ser5 phosphorylation, which mimics early transcription elongation. Using 13C Direct-Detect NMR, we show that the CTD tends to form transient beta strands and beta turns, which are altered differently by Ser2 and Ser5 phosphorylation. Small angle x-ray scattering (SAXS) revealed no significant changes in the CTD global dimensions even at high phosphorylation levels, contradicting the common assumption of phosphorylation-induced chain expansion. Our findings support a transient beta model in which unphosphorylated CTD adopts transient beta strands at Ser2 during transcription pre-initiation. These transient structures are disrupted by Ser5 phosphorylation in early elongation, and later restored by Ser2 phosphorylation near termination for recruiting beta turn-recognizing termination factors.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":"53 1","pages":"108533"},"PeriodicalIF":4.8,"publicationDate":"2025-04-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143876479","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0