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The Arf activator GBF1 localizes to plasma membrane sites involved in cell adhesion and motility. Arf激活因子GBF1定位于参与细胞粘附和运动的质膜部位。
Cellular logistics Pub Date : 2017-03-20 eCollection Date: 2017-01-01 DOI: 10.1080/21592799.2017.1308900
Theodore Busby, Justyna M Meissner, Melanie L Styers, Jay Bhatt, Akhil Kaushik, Anita B Hjelmeland, Elizabeth Sztul
{"title":"The Arf activator GBF1 localizes to plasma membrane sites involved in cell adhesion and motility.","authors":"Theodore Busby, Justyna M Meissner, Melanie L Styers, Jay Bhatt, Akhil Kaushik, Anita B Hjelmeland, Elizabeth Sztul","doi":"10.1080/21592799.2017.1308900","DOIUrl":"https://doi.org/10.1080/21592799.2017.1308900","url":null,"abstract":"GBF1 is ubiquitously expressed in eukaryotic cells and is essential for cellular and organismal life. Depletion of GBF1 from cultured cells induces apoptosis, while mouse GBF1 knockout and D. melan...","PeriodicalId":72547,"journal":{"name":"Cellular logistics","volume":"7 2","pages":"e1308900"},"PeriodicalIF":0.0,"publicationDate":"2017-03-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21592799.2017.1308900","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35163533","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 7
A plasmid library of full-length zebrafish rab proteins for in vivo cell biology. 用于体内细胞生物学的全长斑马鱼蛋白质粒文库。
Cellular logistics Pub Date : 2017-03-01 eCollection Date: 2017-01-01 DOI: 10.1080/21592799.2017.1301151
Thomas E Hall, Nick Martel, Harriet P Lo, Zherui Xiong, Robert G Parton
{"title":"A plasmid library of full-length zebrafish rab proteins for <i>in vivo</i> cell biology.","authors":"Thomas E Hall,&nbsp;Nick Martel,&nbsp;Harriet P Lo,&nbsp;Zherui Xiong,&nbsp;Robert G Parton","doi":"10.1080/21592799.2017.1301151","DOIUrl":"https://doi.org/10.1080/21592799.2017.1301151","url":null,"abstract":"<p><p>The zebrafish is an emerging model for highly sophisticated medium-throughput experiments such as genetic and chemical screens. However, studies of entire protein families within this context are often hampered by poor genetic resources such as clone libraries. Here we describe a complete collection of 76 full-length open reading frame clones for the zebrafish rab protein family. While the mouse genome contains 60 rab genes and the human genome 63, we find that 18 zebrafish rab genes have 2, and in the case of rab38, 3 paralogues. In contrast, we were unable to identify zebrafish orthologues of the mammalian Rab2b, Rab17 or Rab29. We make this resource available through the Addgene repository to facilitate cell biologic approaches using this model.</p>","PeriodicalId":72547,"journal":{"name":"Cellular logistics","volume":"7 1","pages":"e1301151"},"PeriodicalIF":0.0,"publicationDate":"2017-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21592799.2017.1301151","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34902649","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 5
Rab11-FIP1 phosphorylation by MARK2 regulates polarity in MDCK cells. MARK2磷酸化Rab11-FIP1调控MDCK细胞的极性。
Cellular logistics Pub Date : 2017-01-09 eCollection Date: 2017-01-01 DOI: 10.1080/21592799.2016.1271498
Rebecca McRae, Lynne A Lapierre, Elizabeth H Manning, James R Goldenring
{"title":"Rab11-FIP1 phosphorylation by MARK2 regulates polarity in MDCK cells.","authors":"Rebecca McRae,&nbsp;Lynne A Lapierre,&nbsp;Elizabeth H Manning,&nbsp;James R Goldenring","doi":"10.1080/21592799.2016.1271498","DOIUrl":"https://doi.org/10.1080/21592799.2016.1271498","url":null,"abstract":"<p><p>MARK2/Par1b/EMK1, a serine/threonine kinase, is required for correct apical/basolateral membrane polarization in epithelial cells. However, the specific substrates mediating MARK2 action are less well understood. We have now found that MARK2 phosphorylates Rab11-FIP1B/C at serine 234 in a consensus site similar to that previously identified in Rab11-FIP2. In MDCK cells undergoing repolarization after a calcium switch, antibodies specific for pS234-Rab11-FIP1 or pS227-Rab11-FIP2 demonstrate that the spatial and temporal activation of Rab11-FIP1 phosphorylation is distinct from that for Rab11-FIP2. Phosphorylation of Rab11-FIP1 persists through calcium switch and remains high after polarity has been reestablished whereas FIP2 phosphorylation is highest early in reestablishment of polarity but significantly reduced once polarity has been re-established. MARK2 colocalized with FIP1B/C/D and p(S234)-FIP1 <i>in vivo</i>. Overexpression of GFP-Rab11-FIP1C wildtype or non-phosphorylatable GFP-Rab11-FIP1C(S234A) induced two significant phenotypes following calcium switch. Overexpression of FIP1C wildtype and FIP1C(S234A) caused a psuedo-stratification of cells in early time points following calcium switch. At later time points most prominently observed in cells expressing FIP1C(S234A) a significant lateral lumen phenotype was observed, where F-actin-rich lateral lumens appeared demarcated by a ring of ZO1 and also containing ezrin, syntaxin 3 and podocalyxin. In contrast, p120 and E-Cadherin were excluded from the new apical surface at the lateral lumens and now localized to the new lateral surface oriented toward the media. GFP-FIP1C(S234A) localized to membranes deep to the lateral lumens, and immunostaining demonstrated the reorientation of the centrosome and the Golgi apparatus toward the lateral lumen. These results suggest that both Rab11-FIP1B/C and Rab11-FIP2 serve as critical substrates mediating aspects of MARK2 regulation of epithelial polarity.</p>","PeriodicalId":72547,"journal":{"name":"Cellular logistics","volume":"7 1","pages":"e1271498"},"PeriodicalIF":0.0,"publicationDate":"2017-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21592799.2016.1271498","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34902648","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 10
Plasmids for variable expression of proteins targeted to the mitochondrial matrix or intermembrane space 质粒用于针对线粒体基质或膜间隙的蛋白质的可变表达
Cellular logistics Pub Date : 2016-10-01 DOI: 10.1080/21592799.2016.1247939
L. Newman, Cara R. Schiavon, R. Kahn
{"title":"Plasmids for variable expression of proteins targeted to the mitochondrial matrix or intermembrane space","authors":"L. Newman, Cara R. Schiavon, R. Kahn","doi":"10.1080/21592799.2016.1247939","DOIUrl":"https://doi.org/10.1080/21592799.2016.1247939","url":null,"abstract":"ABSTRACT We describe the construction and uses of a series of plasmids for directing expression to varied levels of exogenous proteins targeted to the mitochondrial matrix or intermembrane space. We found that the level of protein expression achieved, the kinetics of expression and mitochondrial import, and half-life after import can each vary with the protein examined. These factors should be considered when directing localization of an exogenous protein to mitochondria for rescue, proteomics, or other approaches. We describe the construction of a collection of plasmids for varied expression of proteins targeted to the mitochondrial matrix or intermembrane space, using previously defined targeting sequences and strength CMV promoters. The limited size of these compartments makes them particularly vulnerable to artifacts from over-expression. We found that different proteins display different kinetics of expression and import that should be considered when analyzing results from this approach. Finally, this collection of plasmids has been deposited in the Addgene plasmid repository to facilitate the ready access and use of these tools.","PeriodicalId":72547,"journal":{"name":"Cellular logistics","volume":"6 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21592799.2016.1247939","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60154778","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 12
Neo1 and phosphatidylethanolamine contribute to vacuole membrane fusion in Saccharomyces cerevisiae Neo1和磷脂酰乙醇胺参与酿酒酵母液泡膜融合
Cellular logistics Pub Date : 2016-07-02 DOI: 10.1080/21592799.2016.1228791
Yuantai Wu, Mehmet Takar, Andrea Cuentas-Condori, T. Graham
{"title":"Neo1 and phosphatidylethanolamine contribute to vacuole membrane fusion in Saccharomyces cerevisiae","authors":"Yuantai Wu, Mehmet Takar, Andrea Cuentas-Condori, T. Graham","doi":"10.1080/21592799.2016.1228791","DOIUrl":"https://doi.org/10.1080/21592799.2016.1228791","url":null,"abstract":"ABSTRACT NEO1 is an essential gene in budding yeast and belongs to a highly conserved subfamily of P-type ATPase genes that encode phospholipid flippases. Inactivation of temperature sensitive neo1ts alleles produces pleiomorphic defects in the secretory and endocytic pathways, including fragmented vacuoles. A screen for multicopy suppressors of neo1-2ts growth defects yielded YPT7, which encodes a Rab7 homolog involved in SNARE-dependent vacuolar fusion. YPT7 suppressed the vacuole fragmentation phenotype of neo1-2, but did not suppress Golgi-associated protein trafficking defects. Neo1 localizes to Golgi and endosomal membranes and was only observed in the vacuole membrane, where Ypt7 localizes, in retromer mutants or when highly overexpressed in wild-type cells. Phosphatidylethanolamine (PE) has been implicated in Ypt7-dependent vacuolar membrane fusion in vitro and is a potential transport substrate of Neo1. Strains deficient in PE synthesis (psd1Δ psd2Δ) displayed fragmented vacuoles and the neo1-2 fragmented vacuole phenotype was also suppressed by overexpression of PSD2, encoding a phosphatidylserine decarboxylase that produces PE at endosomes. In contrast, neo1-2 was not suppressed by overexpression of VPS39, an effector of Ypt7 that forms a membrane contact site potentially involved in PE transfer between vacuoles and mitochondria. These results support the crucial role of PE in vacuole membrane fusion and implicate Neo1 in concentrating PE in the cytosolic leaflet of Golgi and endosomes, and ultimately the vacuole membrane.","PeriodicalId":72547,"journal":{"name":"Cellular logistics","volume":"6 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21592799.2016.1228791","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60155081","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 27
An improved reversibly dimerizing mutant of the FK506-binding protein FKBP 改良的fk506结合蛋白FKBP的可逆二聚化突变体
Cellular logistics Pub Date : 2016-05-16 DOI: 10.1080/21592799.2016.1204848
Juan J. Barrero, E. Papanikou, Jason C. Casler, K. Day, B. Glick
{"title":"An improved reversibly dimerizing mutant of the FK506-binding protein FKBP","authors":"Juan J. Barrero, E. Papanikou, Jason C. Casler, K. Day, B. Glick","doi":"10.1080/21592799.2016.1204848","DOIUrl":"https://doi.org/10.1080/21592799.2016.1204848","url":null,"abstract":"FK506-binding protein (FKBP) is a monomer that binds to FK506, rapamycin, and related ligands. The F36M substitution, in which Phe36 in the ligand-binding pocket is changed to Met, leads to formation of antiparallel FKBP dimers, which can be dissociated into monomers by ligand binding. This FKBP(M) mutant has been employed in the mammalian secretory pathway to generate aggregates that can be dissolved by ligand addition to create cargo waves. However, when testing this approach in yeast, we found that dissolution of FKBP(M) aggregates was inefficient. An improved reversibly dimerizing FKBP formed aggregates that dissolved more readily. This FKBP(L,V) mutant carries the F36L mutation, which increases the affinity of ligand binding, and the I90V mutation, which accelerates ligand-induced dissociation of the dimers. The FKBP(L,V) mutant expands the utility of reversibly dimerizing FKBP.","PeriodicalId":72547,"journal":{"name":"Cellular logistics","volume":"6 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2016-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21592799.2016.1204848","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60154942","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 12
Allosteric properties of PH domains in Arf regulatory proteins. Arf调节蛋白PH结构域的变构特性。
Cellular logistics Pub Date : 2016-04-26 eCollection Date: 2016-04-01 DOI: 10.1080/21592799.2016.1181700
Neeladri Sekhar Roy, Marielle E Yohe, Paul A Randazzo, James M Gruschus
{"title":"Allosteric properties of PH domains in Arf regulatory proteins.","authors":"Neeladri Sekhar Roy,&nbsp;Marielle E Yohe,&nbsp;Paul A Randazzo,&nbsp;James M Gruschus","doi":"10.1080/21592799.2016.1181700","DOIUrl":"https://doi.org/10.1080/21592799.2016.1181700","url":null,"abstract":"<p><p>Pleckstrin Homology (PH) domains bind phospholipids and proteins. They are critical regulatory elements of a number enzymes including guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs) for Ras-superfamily guanine nucleotide binding proteins such as ADP-ribosylation factors (Arfs). Recent studies have indicated that many PH domains may bind more than one ligand cooperatively. Here we discuss the molecular basis of PH domain-dependent allosteric behavior of 2 ADP-ribosylation factor exchange factors, Grp1 and Brag2, cooperative binding of ligands to the PH domains of Grp1 and the Arf GTPase-activating protein, ASAP1, and the consequences for activity of the associated catalytic domains.</p>","PeriodicalId":72547,"journal":{"name":"Cellular logistics","volume":"6 2","pages":"e1181700"},"PeriodicalIF":0.0,"publicationDate":"2016-04-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21592799.2016.1181700","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34462655","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 13
Small GTPases in trafficking - a family approach: Introducing a rolling series focused on groups or families of small GTPases in trafficking. 人口贩运中的小型gtpase -家庭方法:引入滚动系列，重点关注人口贩运中的小型gtpase群体或家庭。
Cellular logistics Pub Date : 2016-04-21 eCollection Date: 2016-01-01 DOI: 10.1080/21592799.2016.1178036
Jennifer L Stow
{"title":"Small GTPases in trafficking - a family approach: Introducing a rolling series focused on groups or families of small GTPases in trafficking.","authors":"Jennifer L Stow","doi":"10.1080/21592799.2016.1178036","DOIUrl":"https://doi.org/10.1080/21592799.2016.1178036","url":null,"abstract":"","PeriodicalId":72547,"journal":{"name":"Cellular logistics","volume":"6 1","pages":"e1178036"},"PeriodicalIF":0.0,"publicationDate":"2016-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21592799.2016.1178036","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34511961","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Degradation elements coincide with cofactor binding sites in a short-lived transcription factor. 降解元件与短寿命转录因子的辅因子结合位点重合。
Cellular logistics Pub Date : 2016-03-08 eCollection Date: 2016-01-01 DOI: 10.1080/21592799.2016.1157664
Christopher M Hickey
{"title":"Degradation elements coincide with cofactor binding sites in a short-lived transcription factor.","authors":"Christopher M Hickey","doi":"10.1080/21592799.2016.1157664","DOIUrl":"https://doi.org/10.1080/21592799.2016.1157664","url":null,"abstract":"Elaborate control of gene expression by transcription factors is common to all kingdoms of life. In eukaryotes, transcription factor abundance and activity are often regulated by targeted proteolysis via the ubiquitin-proteasome system (UPS). The yeast MATα2 (α2) cell type regulator has long served as a model for UPS-dependent transcription factor degradation. Proteolysis of α2 is complex: it involves at least 2 ubiquitylation pathways and multiple regions of α2 affect its degradation. Such complexity also exists for the degradation of other UPS substrates. Here I review α2 degradation, most notably our recent identification of 2 novel degradation elements within α2 that overlap corepressor binding sites. I discuss possible implications of these findings and consider how principles of α2 proteolysis may be relevant to the degradation of other UPS substrates.","PeriodicalId":72547,"journal":{"name":"Cellular logistics","volume":"6 1","pages":"e1157664"},"PeriodicalIF":0.0,"publicationDate":"2016-03-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21592799.2016.1157664","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"34576519","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 8
Plasma membrane regulates Ras signaling networks. 质膜调控Ras信号网络。
Cellular logistics Pub Date : 2016-02-18 eCollection Date: 2015-10-01 DOI: 10.1080/21592799.2015.1136374
Tanmay Sanjeev Chavan, Serena Muratcioglu, Richard Marszalek, Hyunbum Jang, Ozlem Keskin, Attila Gursoy, Ruth Nussinov, Vadim Gaponenko
{"title":"Plasma membrane regulates Ras signaling networks.","authors":"Tanmay Sanjeev Chavan, Serena Muratcioglu, Richard Marszalek, Hyunbum Jang, Ozlem Keskin, Attila Gursoy, Ruth Nussinov, Vadim Gaponenko","doi":"10.1080/21592799.2015.1136374","DOIUrl":"10.1080/21592799.2015.1136374","url":null,"abstract":"<p><p>Ras GTPases activate more than 20 signaling pathways, regulating such essential cellular functions as proliferation, survival, and migration. How Ras proteins control their signaling diversity is still a mystery. Several pieces of evidence suggest that the plasma membrane plays a critical role. Among these are: (1) selective recruitment of Ras and its effectors to particular localities allowing access to Ras regulators and effectors; (2) specific membrane-induced conformational changes promoting Ras functional diversity; and (3) oligomerization of membrane-anchored Ras to recruit and activate Raf. Taken together, the membrane does not only attract and retain Ras but also is a key regulator of Ras signaling. This can already be gleaned from the large variability in the sequences of Ras membrane targeting domains, suggesting that localization, environment and orientation are important factors in optimizing the function of Ras isoforms.</p>","PeriodicalId":72547,"journal":{"name":"Cellular logistics","volume":"5 1","pages":"e1136374"},"PeriodicalIF":0.0,"publicationDate":"2016-02-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/21592799.2015.1136374","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"60154923","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 33
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