Molecular Biology of the Cell最新文献_第4页

Improved tools for live imaging of F-actin structures in yeast. 改进酵母中 F-肌动蛋白结构的实时成像工具。

IF 3.1 3区生物学

Molecular Biology of the Cell Pub Date : 2024-09-01 Epub Date: 2024-07-18 DOI: 10.1091/mbc.E24-05-0212-T

Alison C E Wirshing, Bruce L Goode

{"title":"Improved tools for live imaging of F-actin structures in yeast.","authors":"Alison C E Wirshing, Bruce L Goode","doi":"10.1091/mbc.E24-05-0212-T","DOIUrl":"10.1091/mbc.E24-05-0212-T","url":null,"abstract":"For over 20 years, the most effective probe for live imaging of yeast actin cables has been Abp140-GFP. Here, we report that endogenously-tagged Abp140-GFP poorly decorates actin patches and cables in the bud compartment of yeast cells, while robustly decorating these structures in the mother cell. Using mutagenesis, we found that asymmetric decoration by Abp140 requires F-actin binding. By expressing integrated Bni1-Bnr1 and Bnr1-Bni1 chimeras, we demonstrate that asymmetric cable decoration by Abp140 also does not depend on which formin assembles the cables in each compartment. In contrast, the short actin-binding fragment of Abp140 (known as \"Lifeact\"), fused to 1x or 3xmNeonGreen and expressed from the endogenous ABP140 promoter, uniformly decorates patches and cables in both compartments. Further, this probe dramatically improves live imaging detection of cables (and patches) without altering their in vivo dynamics or cell growth. Improved detection allows us to visualize cables growing inward from the cell cortex and dynamically interacting with the vacuole. This probe also robustly decorates the cytokinetic actomyosin ring. Because Lifeact-3xmNeon expressed at relatively low levels provides intense labeling of cellular F-actin structures, this tool may improve live imaging in other organisms where higher levels of Lifeact expression are detrimental.","PeriodicalId":18735,"journal":{"name":"Molecular Biology of the Cell","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141723950","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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 postdoctoral training program in bioimage analysis. 生物图像分析博士后培训计划。

IF 3.1 3区生物学

Molecular Biology of the Cell Pub Date : 2024-09-01 DOI: 10.1091/mbc.E24-05-0214

Beth A Cimini, Callum Tromans-Coia, David R Stirling, Suganya Sivagurunathan, Rebecca A Senft, Pearl V Ryder, Esteban Miglietta, Paula Llanos, Nasim Jamali, Barbara Diaz-Rohrer, Shatavisha Dasgupta, Mario Cruz, Erin Weisbart, Anne E Carpenter

引用次数: 0

Increased basal fibronectin is sufficient to promote excess endothelial cell matrix assembly causing localized barrier dysfunction. 基础纤连蛋白的增加足以促进内皮细胞基质的过度集结，造成局部屏障功能障碍。

IF 3.1 3区生物学

Molecular Biology of the Cell Pub Date : 2024-09-01 Epub Date: 2024-07-24 DOI: 10.1091/mbc.E24-02-0090

Henry A Resnikoff, Jean E Schwarzbauer

{"title":"Increased basal fibronectin is sufficient to promote excess endothelial cell matrix assembly causing localized barrier dysfunction.","authors":"Henry A Resnikoff, Jean E Schwarzbauer","doi":"10.1091/mbc.E24-02-0090","DOIUrl":"10.1091/mbc.E24-02-0090","url":null,"abstract":"Endothelial cell behavior is regulated by subendothelial extracellular matrix (ECM). The ECM protein fibronectin (FN) is rare in healthy blood vessels but accumulates in disease accompanied by endothelial dysfunctions. Here, we report that excess assembly of FN disrupts key endothelial functions. We mimicked increased FN expression as in diseased stroma by providing exogenous FN basally in a Transwell insert and found dose-dependent upregulation of subendothelial FN matrix assembly. Taking advantage of discontinuous matrix assembly by endothelial cells, we show correlations between regional increases in FN matrix and disruptions in endothelial cell morphology, VE-cadherin junctions, and the cell cycle, all of which were not changed in FN-deficient regions of the monolayer. These changes affected endothelial barrier function with increased monolayer permeability exposing basal regions of high FN matrix and permitting FN-dependent adhesion of MDA-MB-231 tumor cells from the apical side of the monolayer. FN matrix accumulation was quick and increases in FN matrix preceded all other changes in the endothelium. Therefore, subendothelial accumulation of FN matrix is a cause, not an effect, of endothelial monolayer disorganization and leakiness. Regulating FN accumulation in the subendothelial space could be an important target for controlling progression of fibrosis and related diseases.","PeriodicalId":18735,"journal":{"name":"Molecular Biology of the Cell","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141752084","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Caulobacter crescentus RNase E condensation contributes to autoregulation and fitness. 新月芽孢杆菌 RNase E 的凝结有助于自调节和健康。

IF 3.1 3区生物学

Molecular Biology of the Cell Pub Date : 2024-08-01 Epub Date: 2024-06-12 DOI: 10.1091/mbc.E23-12-0493

Vidhyadhar Nandana, Nadra Al-Husini, Arti Vaishnav, Kulathungage H Dilrangi, Jared M Schrader

{"title":"Caulobacter crescentus RNase E condensation contributes to autoregulation and fitness.","authors":"Vidhyadhar Nandana, Nadra Al-Husini, Arti Vaishnav, Kulathungage H Dilrangi, Jared M Schrader","doi":"10.1091/mbc.E23-12-0493","DOIUrl":"10.1091/mbc.E23-12-0493","url":null,"abstract":"RNase E is the most common RNA decay nuclease in bacteria, setting the global mRNA decay rate and scaffolding formation of the RNA degradosome complex and BR-bodies. To properly set the global mRNA decay rate, RNase E from Escherichia coli and neighboring γ-proteobacteria were found to autoregulate RNase E levels via the decay of its mRNA's 5' untranslated region (UTR). While the 5' UTR is absent from other groups of bacteria in the Rfam database, we identified that the α-proteobacterium Caulobacter crescentus RNase E contains a similar 5' UTR structure that promotes RNase E autoregulation. In both bacteria, the C-terminal intrinsically disordered region (IDR) of RNase E is required for proper autoregulation to occur, and this IDR is also necessary and sufficient for RNase E to phase-separate, generating BR-bodies. Using in vitro purified RNase E, we find that the IDR's ability to promote phase separation correlates with enhanced 5' UTR cleavage, suggesting that phase separation of RNase E with the 5' UTR enhances autoregulation. Finally, using growth competition experiments, we find that a strain capable of autoregulation rapidly outcompetes a strain with a 5' UTR mutation that cannot autoregulate, suggesting autoregulation promotes optimal cellular fitness.","PeriodicalId":18735,"journal":{"name":"Molecular Biology of the Cell","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11321048/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141306298","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A genetically-encoded fluorescence-based reporter to spatiotemporally investigate mannose-6-phosphate pathway. 基于基因编码的荧光报告器，用于对 6-磷酸甘露糖途径进行时空研究。

IF 3.1 3区生物学

Molecular Biology of the Cell Pub Date : 2024-08-01 Epub Date: 2024-06-18 DOI: 10.1091/mbc.E23-09-0344

Mallika Bhat, Akshaya Nambiar, Lakshmi Edakkandiyil, Irine Maria Abraham, Ritoprova Sen, Mamta Negi, Ravi Manjithaya

{"title":"A genetically-encoded fluorescence-based reporter to spatiotemporally investigate mannose-6-phosphate pathway.","authors":"Mallika Bhat, Akshaya Nambiar, Lakshmi Edakkandiyil, Irine Maria Abraham, Ritoprova Sen, Mamta Negi, Ravi Manjithaya","doi":"10.1091/mbc.E23-09-0344","DOIUrl":"10.1091/mbc.E23-09-0344","url":null,"abstract":"Maintenance of a pool of active lysosomes with acidic pH and degradative hydrolases is crucial for cell health. Abnormalities in lysosomal function are closely linked to diseases, such as lysosomal storage disorders, neurodegeneration, intracellular infections, and cancer among others. Emerging body of research suggests the malfunction of lysosomal hydrolase trafficking pathway to be a common denominator of several disease pathologies. However, available conventional tools to assess lysosomal hydrolase trafficking are insufficient and fail to provide a comprehensive picture about the trafficking flux and location of lysosomal hydrolases. To address some of the shortcomings, we designed a genetically-encoded fluorescent reporter containing a lysosomal hydrolase tandemly tagged with pH sensitive and insensitive fluorescent proteins, which can spatiotemporally trace the trafficking of lysosomal hydrolases. As a proof of principle, we demonstrate that the reporter can detect perturbations in hydrolase trafficking, that are induced by pharmacological manipulations and pathophysiological conditions like intracellular protein aggregates. This reporter can effectively serve as a probe for mapping the mechanistic intricacies of hydrolase trafficking pathway in health and disease and is a utilitarian tool to identify genetic and pharmacological modulators of this pathway, with potential therapeutic implications.","PeriodicalId":18735,"journal":{"name":"Molecular Biology of the Cell","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11321044/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141419887","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Meiosis-specific functions of kinetochore protein SPC105R required for chromosome segregation in Drosophila oocytes. 果蝇卵母细胞染色体分离所需的动点核蛋白 SPC105R 的减数分裂特异性功能

IF 3.1 3区生物学

Molecular Biology of the Cell Pub Date : 2024-08-01 Epub Date: 2024-06-12 DOI: 10.1091/mbc.E24-02-0067

Jay N Joshi, Neha Changela, Lia Mahal, Janet Jang, Tyler Defosse, Lin-Ing Wang, Arunika Das, Joanatta G Shapiro, Kim McKim

{"title":"Meiosis-specific functions of kinetochore protein SPC105R required for chromosome segregation in Drosophila oocytes.","authors":"Jay N Joshi, Neha Changela, Lia Mahal, Janet Jang, Tyler Defosse, Lin-Ing Wang, Arunika Das, Joanatta G Shapiro, Kim McKim","doi":"10.1091/mbc.E24-02-0067","DOIUrl":"10.1091/mbc.E24-02-0067","url":null,"abstract":"The reductional division of meiosis I requires the separation of chromosome pairs towards opposite poles. We have previously implicated the outer kinetochore protein SPC105R/KNL1 in driving meiosis I chromosome segregation through lateral attachments to microtubules and coorientation of sister centromeres. To identify the domains of SPC105R that are critical for meiotic chromosome segregation, an RNAi-resistant gene expression system was developed. We found that the SPC105R C-terminal domain (aa 1284-1960) is necessary and sufficient for recruiting NDC80 to the kinetochore and building the outer kinetochore. Furthermore, the C-terminal domain recruits BUBR1, which in turn recruits the cohesion protection proteins MEI-S332 and PP2A. Of the remaining 1283 amino acids, we found the first 473 are most important for meiosis. The first 123 amino acids of the N-terminal half of SPC105R contain the conserved SLRK and RISF motifs that are targets of PP1 and Aurora B kinase and are most important for regulating the stability of microtubule attachments and maintaining metaphase I arrest. The region between amino acids 124 and 473 are required for lateral microtubule attachments and biorientation of homologues, which are critical for accurate chromosome segregation in meiosis I.","PeriodicalId":18735,"journal":{"name":"Molecular Biology of the Cell","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11321039/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141306300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Transient PP2A SIP complex localization to mitotic SPBs for SIN inhibition is mediated solely by the Csc1 FHA domain. 瞬时 PP2A SIP 复合物定位到有丝分裂 SPB 以抑制 SIN 的过程完全由 Csc1 FHA 结构域介导。

IF 3.1 3区生物学

Molecular Biology of the Cell Pub Date : 2024-08-01 Epub Date: 2024-06-12 DOI: 10.1091/mbc.E24-04-0196

Alaina H Willet, Liping Ren, Lesley A Turner, Kathleen L Gould

{"title":"Transient PP2A SIP complex localization to mitotic SPBs for SIN inhibition is mediated solely by the Csc1 FHA domain.","authors":"Alaina H Willet, Liping Ren, Lesley A Turner, Kathleen L Gould","doi":"10.1091/mbc.E24-04-0196","DOIUrl":"10.1091/mbc.E24-04-0196","url":null,"abstract":"Many organisms utilize an actin- and myosin-based cytokinetic ring (CR) to help complete cytokinesis. In Schizosaccharomyces pombe, the Septation Initiation Network (SIN) promotes proper CR function and stability. The SIN is a conserved and essential signaling network consisting of a GTPase and a cascade of kinases assembled at the spindle pole body (SPB). The PP2A SIN inhibitory phosphatase (SIP) complex related to the STRIPAK phosphatase complex is one inhibitor of SIN signaling. The SIP consists of Csc1, Csc2, Csc3, Csc4, Paa1, and the phosphatase subunit Ppa3. Here, we determine that the SIP is anchored at the SPB via the Csc1 FHA domain and that constitutive SPB localization of the SIP is lethal due to persistent SIN inhibition. Disrupting SIP docking at the SPB with a point mutation within the FHA domain or eliminating phosphatase activity by introducing a point mutation within Ppa3 resulted in intact SIP complexes without SIN inhibitory function. Lastly, we defined the unique features of Ppa3 that allow it, but not two other PP2A catalytic subunits, to incorporate into the SIP. Overall, we provide insight into how the SIP complex assembles, localizes, and functions to counteract the SIN with spatiotemporal precision during cytokinesis.","PeriodicalId":18735,"journal":{"name":"Molecular Biology of the Cell","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11321038/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141306301","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

An invariant C-terminal tryptophan in McdB mediates its interaction and positioning function with carboxysomes. McdB 中一个不变的 C 端色氨酸介导了它与羧基体的相互作用和定位功能。

IF 3.1 3区生物学

Molecular Biology of the Cell Pub Date : 2024-08-01 Epub Date: 2024-06-26 DOI: 10.1091/mbc.E23-11-0443

Joseph L Basalla, Maria Ghalmi, Y Hoang, Rachel E Dow, Anthony G Vecchiarelli

{"title":"An invariant C-terminal tryptophan in McdB mediates its interaction and positioning function with carboxysomes.","authors":"Joseph L Basalla, Maria Ghalmi, Y Hoang, Rachel E Dow, Anthony G Vecchiarelli","doi":"10.1091/mbc.E23-11-0443","DOIUrl":"10.1091/mbc.E23-11-0443","url":null,"abstract":"Bacterial microcompartments (BMCs) are widespread, protein-based organelles that regulate metabolism. The model for studying BMCs is the carboxysome, which facilitates carbon fixation in several autotrophic bacteria. Carboxysomes can be distinguished as type α or β, which are structurally and phyletically distinct. We recently characterized the maintenance of carboxysome distribution (Mcd) systems responsible for spatially regulating α- and β-carboxysomes, consisting of the proteins McdA and McdB. McdA is an ATPase that drives carboxysome positioning, and McdB is the adaptor protein that directly interacts with carboxysomes to provide cargo specificity. The molecular features of McdB proteins that specify their interactions with carboxysomes, and whether these are similar between α- and β-carboxysomes, remain unknown. Here, we identify C-terminal motifs containing an invariant tryptophan necessary for α- and β-McdBs to associate with α- and β-carboxysomes, respectively. Substituting this tryptophan with other aromatic residues reveals corresponding gradients in the efficiency of carboxysome colocalization and positioning by McdB in vivo. Intriguingly, these gradients also correlate with the ability of McdB to form condensates in vitro. The results reveal a shared mechanism underlying McdB adaptor protein binding to carboxysomes, and potentially other BMCs. Our findings also implicate condensate formation as playing a key role in this association.","PeriodicalId":18735,"journal":{"name":"Molecular Biology of the Cell","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11321042/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141458065","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Simple aneuploidy evades p53 surveillance and promotes niche factor-independent growth in human intestinal organoids. 简单非整倍体可逃避 p53 的监控，并促进人体肠道器官组织中不依赖于生态位因子的生长。

IF 3.1 3区生物学

Molecular Biology of the Cell Pub Date : 2024-08-01 Epub Date: 2024-07-10 DOI: 10.1091/mbc.E24-04-0166

Blake A Johnson, Albert Z Liu, Tianhao Bi, Yi Dong, Taibo Li, Dingjingyu Zhou, Akshay Narkar, Yufei Wu, Sean X Sun, Tatianna C Larman, Jin Zhu, Rong Li

{"title":"Simple aneuploidy evades p53 surveillance and promotes niche factor-independent growth in human intestinal organoids.","authors":"Blake A Johnson, Albert Z Liu, Tianhao Bi, Yi Dong, Taibo Li, Dingjingyu Zhou, Akshay Narkar, Yufei Wu, Sean X Sun, Tatianna C Larman, Jin Zhu, Rong Li","doi":"10.1091/mbc.E24-04-0166","DOIUrl":"10.1091/mbc.E24-04-0166","url":null,"abstract":"Aneuploidy is nearly ubiquitous in tumor genomes, but the role of aneuploidy in the early stages of cancer evolution remains unclear. Here, by inducing heterogeneous aneuploidy in non-transformed human colon organoids (colonoids), we investigated how the effects of aneuploidy on cell growth and differentiation may promote malignant transformation. Previous work implicated p53 activation as a downstream response to aneuploidy induction. We found that simple aneuploidy, characterized by 1-3 gained or lost chromosomes, resulted in little or modest p53 activation and cell cycle arrest when compared with more complex aneuploid cells. Single-cell RNA sequencing analysis revealed that the degree of p53 activation was strongly correlated with karyotype complexity. Single-cell tracking showed that cells could continue to divide despite the observation of one to a few lagging chromosomes. Unexpectedly, colonoids with simple aneuploidy exhibited impaired differentiation after niche factor withdrawal. These findings demonstrate that simple aneuploid cells can escape p53 surveillance and may contribute to niche factor-independent growth of cancer-initiating colon stem cells.","PeriodicalId":18735,"journal":{"name":"Molecular Biology of the Cell","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11321050/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141580243","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The core spindle pole body scaffold Ppc89 links the pericentrin orthologue Pcp1 to the fission yeast spindle pole body via an evolutionarily conserved interface. 核心纺锤极体支架Ppc89通过一个进化保守的界面将包心蛋白直向同源物Pcp1与裂殖酵母纺锤极体连接起来。

IF 3.1 3区生物学

Molecular Biology of the Cell Pub Date : 2024-08-01 Epub Date: 2024-07-10 DOI: 10.1091/mbc.E24-05-0220

Jun-Song Chen, Maya G Igarashi, Liping Ren, Sarah M Hanna, Lesley A Turner, Nathan A McDonald, Janel R Beckley, Alaina H Willet, Kathleen L Gould

{"title":"The core spindle pole body scaffold Ppc89 links the pericentrin orthologue Pcp1 to the fission yeast spindle pole body via an evolutionarily conserved interface.","authors":"Jun-Song Chen, Maya G Igarashi, Liping Ren, Sarah M Hanna, Lesley A Turner, Nathan A McDonald, Janel R Beckley, Alaina H Willet, Kathleen L Gould","doi":"10.1091/mbc.E24-05-0220","DOIUrl":"10.1091/mbc.E24-05-0220","url":null,"abstract":"Centrosomes and spindle pole bodies (SPBs) are important for mitotic spindle formation and serve as cellular signaling platforms. Although centrosomes and SPBs differ in morphology, many mechanistic insights into centrosome function have been gleaned from SPB studies. In the fission yeast Schizosaccharomyces pombe, the α-helical protein Ppc89, identified based on its interaction with the septation initiation network scaffold Sid4, comprises the SPB core. High-resolution imaging has suggested that SPB proteins assemble on the Ppc89 core during SPB duplication, but such interactions are undefined. Here, we define a connection between Ppc89 and the essential pericentrin Pcp1. Specifically, we found that a predicted third helix within Ppc89 binds the Pcp1 pericentrin-AKAP450 centrosomal targeting (PACT) domain complexed with calmodulin. Ppc89 helix 3 contains similarity to present in the N-terminus of Cep57 (PINC) motifs found in the centrosomal proteins fly SAS-6 and human Cep57 and also to the S. cerevisiae SPB protein Spc42. These motifs bind pericentrin-calmodulin complexes and AlphaFold2 models suggest a homologous complex assembles in all four organisms. Mutational analysis of the S. pombe complex supports the importance of Ppc89-Pcp1 binding interface in vivo. Our studies provide insight into the core architecture of the S. pombe SPB and suggest an evolutionarily conserved mechanism of scaffolding pericentrin-calmodulin complexes for mitotic spindle formation.","PeriodicalId":18735,"journal":{"name":"Molecular Biology of the Cell","volume":null,"pages":null},"PeriodicalIF":3.1,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11321043/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141580245","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0