Journal of Biological Chemistry最新文献_第7页

In vitro reconstitution reveals substrate selectivity of protein S-acyltransferases.

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-03-14 DOI: 10.1016/j.jbc.2025.108406

Tanmay Mondal, James Song, Anirban Banerjee

{"title":"In vitro reconstitution reveals substrate selectivity of protein S-acyltransferases.","authors":"Tanmay Mondal, James Song, Anirban Banerjee","doi":"10.1016/j.jbc.2025.108406","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108406","url":null,"abstract":"Protein S-acylation, commonly known as protein palmitoylation, is the most prevalent form of protein lipidation with ∼6000 target proteins and in humans, is catalyzed by 23 integral membrane enzymes of the zDHHC family. Recognition of its importance in cellular physiology as well as human diseases has undergone an explosive growth in recent years. Yet, the nature of zDHHC-substrate interactions has remained poorly understood for most zDHHC enzymes. Cell-based experiments indicate a promiscuous and complex zDHHC-substrate network whereas lack of in vitro reconstitution experiments has impeded insights into the nature of discrete zDHHC-substrate interactions. Here we report a substrate S-acylation reconstitution assay, called the Pep-PAT assay, using purified enzyme and peptide fragments of substrates. We use the Pep-PAT assay to investigate the substrate S-acylation of three different zDHHC enzymes on seven different substrates. Remarkably, all the zDHHC enzymes showed robust activity with certain substrates but not others. These in vitro reconstitution experiments indicate that there is a preferred substrate hierarchy for zDHHC enzymes. We further used the Pep-PAT assay to interrogate the role of neighboring residues around the target cysteine on S-acylation of PSD-95 and SARS-CoV-2 Spike protein. Select residues around the target cysteines have distinct impact on substrate S-acylation, leading to the first insights into how neighboring residues around the target cysteine affect substrate S-acylation by zDHHC enzymes. Finally, we validated the impact of neighboring residues on substrate S-acylation using in cellulo assays. Our experiments build a framework for understanding substrate S-acylation by zDHHC enzymes.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"108406"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143639543","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

Gonadotropin-releasing hormone regulates transcription of the inhibin B co-receptor, TGFBR3L, via early growth response 1. 促性腺激素释放激素通过早期生长应答 1 调节抑制素 B 共受体 TGFBR3L 的转录。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-03-14 DOI: 10.1016/j.jbc.2025.108405

Yeu-Farn Lin, Evan R S Buddle, Hailey Schultz, Xiang Zhou, Luisina Ongaro, Mary Loka, Carlos A I Alonso, Ulrich Boehm, Raj Duggavathi, Daniel J Bernard

{"title":"Gonadotropin-releasing hormone regulates transcription of the inhibin B co-receptor, TGFBR3L, via early growth response 1.","authors":"Yeu-Farn Lin, Evan R S Buddle, Hailey Schultz, Xiang Zhou, Luisina Ongaro, Mary Loka, Carlos A I Alonso, Ulrich Boehm, Raj Duggavathi, Daniel J Bernard","doi":"10.1016/j.jbc.2025.108405","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108405","url":null,"abstract":"Follicle-stimulating hormone (FSH), a product of pituitary gonadotrope cells, regulates gonadal function and fertility. FSH production is stimulated by gonadotropin-releasing hormone (GnRH) and activin-class ligands of the TGFβ family. Inhibin A and B are TGFβ proteins that suppress FSH synthesis by competitively binding activin type II receptors in concert with the co-receptors betaglycan (TGFBR3) and TGFBR3L. Betaglycan mediates the actions of both inhibins and is broadly expressed. In contrast, TGFBR3L is inhibin B-specific and selectively expressed in gonadotropes. This cell-restricted expression is driven, in part, by steroidogenic factor 1 (SF-1, NR5A1), which stimulates Tgfbr3l/TGFBR3L transcription via two conserved promoter elements. Tgfbr3l expression is lost in mice lacking SF-1 in gonadotropes. However, SF-1 alone is unlikely to fully explain gonadotrope-restricted Tgfbr3l/TGFBR3L expression. Here, we report that GnRH induces binding of the transcription factor, early growth response 1 (EGR1), to the murine Tgfbr3l and human TGFBR3L promoters at a conserved cis-element between the two SF-1 binding sites. In homologous LβT2 cells, GnRH stimulation of Tgfbr3l/TGFBR3L promoter-reporters depends on EGR1 binding to this cis-element. In heterologous cells, over-expressed EGR1 independently and synergistically with SF-1 activates Tgfbr3l/TGFBR3L promoter-reporter activities. In vivo, Tgfbr3l mRNA expression is reduced in pituitaries of 1) GnRH-deficient mice, 2) wild-type mice treated with a GnRH receptor antagonist, and 3) gonadotrope-specific Egr1 knockout mice. Gonadectomy, which increases GnRH pulse frequency, enhances Tgfbr3l expression in control but not gonadotrope-specific Egr1 knockouts. Collectively, these data indicate that GnRH stimulates Tgfbr3l/TGFBR3L transcription via EGR1, which acts with SF-1 through conserved promoter elements.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"108405"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143639500","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

A conserved phosphorylation mechanism for regulating the interaction between the CMG replicative helicase and its forked DNA substrate.

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-03-14 DOI: 10.1016/j.jbc.2025.108408

Sandra Koit, Nele Tamberg, Allan Reinapae, Lauri Peil, Arnold Kristjuhan, Ivar Ilves

{"title":"A conserved phosphorylation mechanism for regulating the interaction between the CMG replicative helicase and its forked DNA substrate.","authors":"Sandra Koit, Nele Tamberg, Allan Reinapae, Lauri Peil, Arnold Kristjuhan, Ivar Ilves","doi":"10.1016/j.jbc.2025.108408","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108408","url":null,"abstract":"The CMG helicase is a crucial enzyme complex that plays a vital role in the replication of genomic DNA in eukaryotes. Besides unwinding the DNA template and coordinating the replisome's structure, it is also a key target for signaling pathways that regulate the replication process. We show that a specific serine/threonine residue in the MCM3 subunit of CMG, which has been previously linked to phosphorylation-dependent control mechanisms of genomic DNA replication in human cells, is a conserved phosphorylation site for Chk1 and potentially other protein kinases. This suggests a conserved regulatory mechanism associated with it in metazoans and several other eukaryotes, including budding yeast. Our in vitro analysis links this mechanism directly to the modulation of the CMG helicase activity by impacting its interactions with the forked DNA substrate. Further supporting its conserved role in regulation, we found that phosphomimetic substitution with aspartic acid and alanine knock-out of this conserved residue lead to opposite phenotypic defects in the growth of budding yeast cells. These findings outline a candidate conserved phosphorylation pathway for regulating genomic DNA replication in eukaryotes, which adjusts the interactions between the replicative helicase complex and its DNA substrate according to the specific needs of various physiological conditions.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"108408"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143639457","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 Vitro Inhibition of Voltage-Dependent Sodium Currents by the Antifungal Drug Amorolfine.

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-03-14 DOI: 10.1016/j.jbc.2025.108407

Mohammad-Reza Ghovanloo, Sidharth Tyagi, Philip R Effraim, Stephen G Waxman

{"title":"In Vitro Inhibition of Voltage-Dependent Sodium Currents by the Antifungal Drug Amorolfine.","authors":"Mohammad-Reza Ghovanloo, Sidharth Tyagi, Philip R Effraim, Stephen G Waxman","doi":"10.1016/j.jbc.2025.108407","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108407","url":null,"abstract":"Voltage-gated sodium (Nav) channels are critical for electrical signaling, and their pharmacological modulation can be leveraged for the development of therapeutic agents targeting various disorders. The local anesthetic (LA) site on Nav channels is particularly important, as it is a common target for many clinically used inhibitors, including anticonvulsants and antiarrhythmics. Our goal was to identify novel Nav channel inhibitors by leveraging physicochemical criteria, focusing on potential LA site binding candidates. We identified amorolfine (AMF), a phenyl-propyl morpholine derivative, as a putative modulator. Our results demonstrate that AMF acts as a state-dependent inhibitor of Nav channels, with a ∼30-fold preference for inactivated states. It stabilizes channel inactivation and prevents channel from conducting, driven through its stabilization of inactivation. These findings suggest that AMF represents a new compound that inhibits Nav channels, offering insights into the development of future therapeutic agents targeting Nav and potentially other ion channels.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"108407"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143639578","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

Pak1 dysregulates Pyruvate metabolism in PDAC cells by exerting a phosphorylation-mediated regulatory effect on PDHA1. Pak1 通过对 PDHA1 发挥磷酸化介导的调控作用，使 PDAC 细胞的丙酮酸代谢失调。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-03-14 DOI: 10.1016/j.jbc.2025.108409

Sowmiya Murugan, Srikanth Swamy Swaroop B, Prarthana Gopinath, Roshni Saravanan, Sandhya Sundaram, Gouthaman Shanmugasundaram, Ganesh Venkatraman, Suresh Kumar Rayala

{"title":"Pak1 dysregulates Pyruvate metabolism in PDAC cells by exerting a phosphorylation-mediated regulatory effect on PDHA1.","authors":"Sowmiya Murugan, Srikanth Swamy Swaroop B, Prarthana Gopinath, Roshni Saravanan, Sandhya Sundaram, Gouthaman Shanmugasundaram, Ganesh Venkatraman, Suresh Kumar Rayala","doi":"10.1016/j.jbc.2025.108409","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108409","url":null,"abstract":"Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive form of pancreatic cancer with the worst prognosis. Treating PDAC poses significant challenges, as tumor cells adapt metabolic alterations to thrive in the hypoxic environment created by desmoplasia surrounding the tumor cells. p21-activated kinase (Pak1), a serine-threonine kinase is found to be upregulated in many solid tumors and promotes tumor progression via diverse signalling pathways. In this study, we focussed on exploring the role of Pak1 in mediating tumor cell metabolism. Deletion of the Pak1 gene reduced the tumorigenic potential of PDAC cells. Also, Pak1 regulated both glycolysis and mitochondrial respiration in PDAC cells, contributing to the Warburg phenomenon. Untargeted metabolomic analysis revealed that Pak1 was strongly associated with Pyruvate metabolism. Interestingly, we found that Pak1 interacted and phosphorylated Pyruvate dehydrogenase E1α (PDHA1) at Serine 152. This phosphorylation negatively regulates PDHA1 activity, implying the direct regulatory role of Pak1 in Pyruvate metabolism. Moreover, deleting the Pak1 gene altered the expression and activity of PDHA1 and LDHA, as both are involved in regulating the direction of pyruvate flux inside the cells. Our study demonstrated that Pak1 plays a significant role in PDAC metabolism and Warburg effect, partly by phosphorylating PDHA1.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"108409"},"PeriodicalIF":4.0,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143639553","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

Characterization of quinazolinone calcilytic therapy for autosomal dominant hypocalcemia type 1 (ADH1).

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-03-12 DOI: 10.1016/j.jbc.2025.108404

Fadil M Hannan, Kreepa G Kooblall, Mark Stevenson, Taha Elajnaf, Fangyu Liu, Kate E Lines, Xin Meng, Michelle Stewart, Sara Wells, Edward F Nemeth, Brian K Shoichet, Michaela Kneissel, Juerg A Gasser, Rajesh V Thakker

{"title":"Characterization of quinazolinone calcilytic therapy for autosomal dominant hypocalcemia type 1 (ADH1).","authors":"Fadil M Hannan, Kreepa G Kooblall, Mark Stevenson, Taha Elajnaf, Fangyu Liu, Kate E Lines, Xin Meng, Michelle Stewart, Sara Wells, Edward F Nemeth, Brian K Shoichet, Michaela Kneissel, Juerg A Gasser, Rajesh V Thakker","doi":"10.1016/j.jbc.2025.108404","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108404","url":null,"abstract":"Germline gain-of-function mutations of the calcium-sensing receptor (CaSR) result in autosomal dominant hypocalcemia type 1 (ADH1), which may cause symptomatic hypocalcemia with low parathyroid hormone (PTH) concentrations. Negative allosteric CaSR modulators, known as calcilytics, have potential as a targeted ADH1 therapy and consist of two main classes, which are the amino alcohols and the quinazolinones. Amino alcohol calcilytics have been extensively assessed as ADH1 therapies, but may not be effective for all ADH1-causing mutations. We therefore conducted in silico, in vitro and in vivo evaluations of quinazolinone calcilytics (ATF936 and AXT914) as an alternate ADH1 treatment. Calcilytic docking studies were performed using reported cryo-electron microscopy CaSR structures. In vitro dose-response studies were performed using CaSR-expressing HEK293 cells and in vivo studies undertaken in mice with a gain-of-function CaSR mutation, Leu723Gln, known as Nuf. ATF936 and AXT914, as well as the amino alcohol calcilytics, NPS 2143 and NPSP795, were shown to bind at a common region with the CaSR transmembrane domain, which is also an ADH1 mutational hotspot. Treatment of cells expressing the Nuf mutant (Gln723) CaSR with 1-20nM AXT914 caused dose-dependent decreases in CaSR-mediated intracellular calcium responses with 10nM AXT914 normalising the gain-of-function caused by the mutant CaSR. Oral administration of 10 mg/kg AXT914 to Nuf mice increased plasma PTH to 104±29 pmol/L compared to 23±4 pmol/L for vehicle-treated mice, p<0.05; and increased plasma albumin-adjusted calcium to 2.03±0.02 mmol/L compared to 1.84±0.02 mmol/L for vehicle-treated mice, p<0.001. These studies indicate that the quinazolinone calcilytics may have potential for treating ADH1.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"108404"},"PeriodicalIF":4.0,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143630493","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 E3 ubiquitin ligase RNF126 facilitates quality control of unimported mitochondrial membrane proteins.

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-03-12 DOI: 10.1016/j.jbc.2025.108403

Di Liu, Xin-Yu Huo, Xiaoli Zhang, Zai-Rong Zhang

{"title":"The E3 ubiquitin ligase RNF126 facilitates quality control of unimported mitochondrial membrane proteins.","authors":"Di Liu, Xin-Yu Huo, Xiaoli Zhang, Zai-Rong Zhang","doi":"10.1016/j.jbc.2025.108403","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108403","url":null,"abstract":"Pathological stress can lead to failure in the translocation of mitochondrial proteins, resulting in accumulation of unimported proteins within the cytosol and upregulation of proteasome for their quality control. Malfunction or delay in protein clearance causes dysregulation of mitochondrial protein homeostasis, cellular toxicity, and diseases. Ubiquilins (UBQLNs) are known to serve as chaperone which associates with unimported mitochondrial membrane protein precursors, and facilitates their proteasomal degradation. However, how UBQLN-engaged proteins are ubiquitinated and efficiently targeted to the proteasome are poorly understood. Here, using mitochondrial membrane protein ATP5G1 as a model substrate, we report that E3 ubiquitin ligase RNF126 interacts with substrate-engaged UBQLN1, thereby promoting ubiquitination and degradation of unimported proteins during mitochondrial stress. We find that UBQLN1's ubiquitin-associated domain (UBA) recruits RNF126 when its middle domain binds to unimported protein substrate. Recombinant RNF126 forms ternary complex with UBQLN1 and pATP5G1 in vitro and catalyzes ubiquitination of UBQLN1-bound ATP5G1. Without RNF126, proteasomal degradation of ATP5G1 was compromised. These results explain how RNF126 and ubiquilins interplay to ensure specific quality control of unimported mitochondrial membrane proteins under pathophysiological conditions.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"108403"},"PeriodicalIF":4.0,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143630495","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

Characterization of a Rhodopsin-Phosphodiesterase from Choanoeca flexa to be combined with Rhodopsin-Cyclases for bidirectional optogenetic cGMP control.

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-03-11 DOI: 10.1016/j.jbc.2025.108401

Nicolas Liem, Anika Spreen, Arita Silapētere, Peter Hegemann

{"title":"Characterization of a Rhodopsin-Phosphodiesterase from Choanoeca flexa to be combined with Rhodopsin-Cyclases for bidirectional optogenetic cGMP control.","authors":"Nicolas Liem, Anika Spreen, Arita Silapētere, Peter Hegemann","doi":"10.1016/j.jbc.2025.108401","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108401","url":null,"abstract":"Rhodopsin phosphodiesterases (RhPDEs) were first discovered in the choanoflagellate Salpingoeca rosetta, but their physiological role remained unknown. Their light-dependent modulation was found to be low, limiting optogenetic application. However, recent in vivo studies in the choanoflagellate Choanoeca flexa revealed a strong linkage of RhPDE to the actomyosin-mediated contraction and colony sheet inversion and identified downstream cGMP effectors. Through screening various RhPDE variants from C.flexa, we identified four photomodulated PDEs of which CfRhPDE1 revealed the highest cGMP affinity and the most pronounced light regulation with Km values of 1.9 and 4.4 μM in light and darkness. By co-expressing CfRhPDE1 with the Rhodopsin-guanylyl-cyclase from the fungus Catenaria anguillulae and a cyclic nucleotide-gated ion channel from olfactory neurons in ND7/23 cells, we demonstrate bidirectional dual-color modulation of cGMP levels and ion channel conductance. Together with spectroscopic characterization, our fast functional recordings suggest that the M-state of the photocycle initiates functional changes in the phosphodiesterase domain via rapid rhodopsin-PDE coupling. With efficient expression and 3.5 s lifetime of the active state, this protein provides high photosensitivity to the host cells. This demonstrates that RhPDEs can regulate cGMP signaling in mammalian cells on a subsecond timescale, closing a present gap in optogenetics and assisting researchers in setting up multicomponent optogenetic systems for bidirectional control of cyclic nucleotides.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"108401"},"PeriodicalIF":4.0,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143624784","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

Declining activity of serum response factor in aging aorta in relation to aneurysm progression.

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-03-11 DOI: 10.1016/j.jbc.2025.108400

Catarina Rippe, Joakim Armstrong Bastrup, Johan Holmberg, Katarzyna Kawka, Marycarmen Arévalo Martinez, Sebastian Albinsson, Thomas A Jepps, Karl Swärd

{"title":"Declining activity of serum response factor in aging aorta in relation to aneurysm progression.","authors":"Catarina Rippe, Joakim Armstrong Bastrup, Johan Holmberg, Katarzyna Kawka, Marycarmen Arévalo Martinez, Sebastian Albinsson, Thomas A Jepps, Karl Swärd","doi":"10.1016/j.jbc.2025.108400","DOIUrl":"https://doi.org/10.1016/j.jbc.2025.108400","url":null,"abstract":"Age is a critical determinant of arterial disease, including aneurysm formation. Here, to understand the impact of aging on the arterial transcriptome, we leveraged RNA-sequencing data to define transcripts that change with advancing age in human arteries. Among the most repressed transcripts in aged individuals were those that are relevant for actomyosin structure and organization, including both myosin light chain kinase (MYLK) and smooth muscle γ-actin (ACTG2). This was associated with a reduction of serum response factor (SRF), which controls these transcripts via defined promoter elements. To determine the consequences of isolated Srf depletion, we conditionally deleted Srf in vascular smooth muscle of young mice (i8-SRF-KO mice). This led to a reduction of the SRF regulon, including Mylk and Actg2, and impaired arterial contractility, but left endothelial-dependent dilatation unaffected. Srf-depletion also increased aortic diameter and Alcian blue staining of the aortic media, which are cardinal features of aortopathy, such as aortic aneurysmal disease. Despite this, i8-SRF-KO mice were protected from aortic lesions elicited by angiotensin II (AngII). Proteomics demonstrated that Srf-depletion mimicked a protein signature of AngII treatment involving increases of the mechanoresponsive transcriptional co-activators YAP and TAZ and reduction of the Hippo kinase Lats2. Protection from aortopathy could be overcome by changing the order of knockout induction and AngII administration resulting in advanced aneurysms in both i8-SRF-KO and control mice. Our work provides important insights into the molecular underpinnings of age-dependent changes in aortic function and mechanisms of adaptation in hypertension.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"108400"},"PeriodicalIF":4.0,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143624785","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

YibN, a bona fide interactor of the bacterial YidC insertase with effects on membrane protein insertion and membrane lipid production. YibN是细菌YidC插入酶的一个真正的相互作用者，对膜蛋白插入和膜脂生成有影响。

IF 4 2区生物学

Journal of Biological Chemistry Pub Date : 2025-03-11 DOI: 10.1016/j.jbc.2025.108395

Zhiyu Zhao, Nachi Yamamoto, John W Young, Nestor Solis, Amos Fong, Mohammed Al-Seragi, Sungyoung Kim, Hiroyuki Aoki, Sadhna Phanse, Hai-Tuong Le, Christopher M Overall, Hanako Nishikawa, Mohan Babu, Ken-Ichi Nishiyama, Franck Duong van Hoa

{"title":"YibN, a bona fide interactor of the bacterial YidC insertase with effects on membrane protein insertion and membrane lipid production.","authors":"Zhiyu Zhao, Nachi Yamamoto, John W Young, Nestor Solis, Amos Fong, Mohammed Al-Seragi, Sungyoung Kim, Hiroyuki Aoki, Sadhna Phanse, Hai-Tuong Le, Christopher M Overall, Hanako Nishikawa, Mohan Babu, Ken-Ichi Nishiyama, Franck Duong van Hoa","doi":"10.1016/j.jbc.2025.108395","DOIUrl":"10.1016/j.jbc.2025.108395","url":null,"abstract":"YidC, a prominent member of the Oxa1 superfamily, is essential for the biogenesis of the bacterial inner membrane, significantly influencing its protein composition and lipid organization. It interacts with the Sec translocon, aiding the proper folding of multi-pass membrane proteins. It also functions independently, serving as an insertase and lipid scramblase, augmenting the insertion of smaller membrane proteins while contributing to the organization of the bilayer. Despite the wealth of structural and biochemical data available, how YidC operates remains unclear. To investigate this, we employed proximity-dependent biotin labeling (BioID) in Escherichia coli, leading to the identification of YibN as a crucial component within the YidC protein environment. We then demonstrated the association between YidC and YibN by affinity purification-mass spectrometry assays conducted on native membranes, with further confirmation using on-gel binding assays with purified proteins. Co-expression studies and in vitro assays indicated that YibN enhances the production and membrane insertion of YidC substrates, such as M13 and Pf3 phage coat proteins, ATP synthase subunit c, and various small membrane proteins like SecG. Additionally, the overproduction of YibN was found to stimulate membrane lipid production and promote inner membrane proliferation, perhaps by interfering with YidC lipid scramblase activity. Consequently, YibN emerges as a significant physical and functional interactor of YidC, influencing membrane protein insertion and lipid organization.","PeriodicalId":15140,"journal":{"name":"Journal of Biological Chemistry","volume":" ","pages":"108395"},"PeriodicalIF":4.0,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143624713","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