Eukaryotic Cell最新文献_第10页

Article of Significant Interest Selected from This Issue by the Editors 由编辑从本期杂志中选出的具有重要意义的文章

Eukaryotic Cell Pub Date : 2015-04-01 DOI: 10.1128/ec.00034-15

引用次数: 0

How the necrotrophic fungus Alternaria brassicicola kills plant cells remains an enigma. 坏死性真菌十字花菌是如何杀死植物细胞的仍然是个谜。

Eukaryotic Cell Pub Date : 2015-04-01 Epub Date: 2015-02-13 DOI: 10.1128/EC.00226-14

Yangrae Cho

{"title":"How the necrotrophic fungus Alternaria brassicicola kills plant cells remains an enigma.","authors":"Yangrae Cho","doi":"10.1128/EC.00226-14","DOIUrl":"https://doi.org/10.1128/EC.00226-14","url":null,"abstract":"Alternaria species are mainly saprophytic fungi, but some are plant pathogens. Seven pathotypes of Alternaria alternata use secondary metabolites of host-specific toxins as pathogenicity factors. These toxins kill host cells prior to colonization. Genes associated with toxin synthesis reside on conditionally dispensable chromosomes, supporting the notion that pathogenicity might have been acquired several times by A. alternata. Alternaria brassicicola, however, seems to employ a different mechanism. Evidence on the use of host-specific toxins as pathogenicity factors remains tenuous, even after a diligent search aided by full-genome sequencing and efficient reverse-genetics approaches. Similarly, no individual genes encoding lipases or cell wall-degrading enzymes have been identified as strong virulence factors, although these enzymes have been considered important for fungal pathogenesis. This review describes our current understanding of toxins, lipases, and cell wall-degrading enzymes and their roles in the pathogenesis of A. brassicicola compared to those of other pathogenic fungi. It also describes a set of genes that affect pathogenesis in A. brassicicola. They are involved in various cellular functions that are likely important in most organisms and probably indirectly associated with pathogenesis. Deletion or disruption of these genes results in weakly virulent strains that appear to be sensitive to the defense mechanisms of host plants. Finally, this review discusses the implications of a recent discovery of three important transcription factors associated with pathogenesis and the putative downstream genes that they regulate. ","PeriodicalId":11891,"journal":{"name":"Eukaryotic Cell","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2015-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1128/EC.00226-14","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"33053619","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: 45

A fungal sarcolemmal membrane-associated protein (SLMAP) homolog plays a fundamental role in development and localizes to the nuclear envelope, endoplasmic reticulum, and mitochondria. 真菌肌层膜相关蛋白(SLMAP)同源物在发育过程中起着重要作用，定位于核膜、内质网和线粒体。

Eukaryotic Cell Pub Date : 2015-04-01 Epub Date: 2014-12-19 DOI: 10.1128/EC.00241-14

Steffen Nordzieke, Thomas Zobel, Benjamin Fränzel, Dirk A Wolters, Ulrich Kück, Ines Teichert

{"title":"A fungal sarcolemmal membrane-associated protein (SLMAP) homolog plays a fundamental role in development and localizes to the nuclear envelope, endoplasmic reticulum, and mitochondria.","authors":"Steffen Nordzieke, Thomas Zobel, Benjamin Fränzel, Dirk A Wolters, Ulrich Kück, Ines Teichert","doi":"10.1128/EC.00241-14","DOIUrl":"https://doi.org/10.1128/EC.00241-14","url":null,"abstract":"Sarcolemmal membrane-associated protein (SLMAP) is a tail-anchored protein involved in fundamental cellular processes, such as myoblast fusion, cell cycle progression, and chromosomal inheritance. Further, SLMAP misexpression is associated with endothelial dysfunctions in diabetes and cancer. SLMAP is part of the conserved striatin-interacting phosphatase and kinase (STRIPAK) complex required for specific signaling pathways in yeasts, filamentous fungi, insects, and mammals. In filamentous fungi, STRIPAK was initially discovered in Sordaria macrospora, a model system for fungal differentiation. Here, we functionally characterize the STRIPAK subunit PRO45, a homolog of human SLMAP. We show that PRO45 is required for sexual propagation and cell-to-cell fusion and that its forkhead-associated (FHA) domain is essential for these processes. Protein-protein interaction studies revealed that PRO45 binds to STRIPAK subunits PRO11 and SmMOB3, which are also required for sexual propagation. Superresolution structured-illumination microscopy (SIM) further established that PRO45 localizes to the nuclear envelope, endoplasmic reticulum, and mitochondria. SIM also showed that localization to the nuclear envelope requires STRIPAK subunits PRO11 and PRO22, whereas for mitochondria it does not. Taken together, our study provides important insights into fundamental roles of the fungal SLMAP homolog PRO45 and suggests STRIPAK-related and STRIPAK-unrelated functions. ","PeriodicalId":11891,"journal":{"name":"Eukaryotic Cell","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2015-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1128/EC.00241-14","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32922175","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: 50

Role of Tec1 in the development, architecture, and integrity of sexual biofilms of Candida albicans. Tec1在白色念珠菌性生物膜的发育、结构和完整性中的作用。

Eukaryotic Cell Pub Date : 2015-03-01 Epub Date: 2015-01-02 DOI: 10.1128/EC.00224-14

Karla J Daniels, Thyagarajan Srikantha, Claude Pujol, Yang-Nim Park, David R Soll

{"title":"Role of Tec1 in the development, architecture, and integrity of sexual biofilms of Candida albicans.","authors":"Karla J Daniels, Thyagarajan Srikantha, Claude Pujol, Yang-Nim Park, David R Soll","doi":"10.1128/EC.00224-14","DOIUrl":"https://doi.org/10.1128/EC.00224-14","url":null,"abstract":"MTL-homozygous ( A: / A: or α/α) white cells form a complex sexual biofilm that exhibits the same architecture as that of MTL-heterozygous ( A: /α) pathogenic biofilms. However, the former is regulated by the mitogen-activated protein (MAP) kinase pathway, while the latter is regulated by the Ras1/cyclic AMP (cAMP) pathway. We previously demonstrated that in the formation of an MTL-homozygous, mature (48 h) sexual biofilm in RPMI 1640 medium, the MAP kinase pathway targets Tec1 rather than Cph1, the latter of which is the target of the same pathway, but for the opaque cell mating response. Here we continued our analysis of the role of Tec1 by comparing the effects of deleting TEC1 on initial adhesion to silicone elastomer, high-resolution confocal microscopy assessments of the stages and cellular phenotypes during the 48 h of biofilm development, human white cell penetration, and biofilm fragility. We show that although Tec1 plays only a minor role in initial adhesion to the silicone elastomer, it does play a major role in the growth of the basal yeast cell polylayer, vertical extension of hyphae and matrix deposition in the upper portion of the biofilm, final biofilm thickness, penetrability of human white blood cells, and final biofilm integrity (i.e., resistance to fluid flow). These results provide a more detailed description of normal biofilm development and architecture and confirm the central role played by the transcription factor Tec1 in the biofilm model employed here. ","PeriodicalId":11891,"journal":{"name":"Eukaryotic Cell","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2015-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1128/EC.00224-14","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32947279","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: 19

Dynamics of the lipid droplet proteome of the Oleaginous yeast rhodosporidium toruloides. 产油酵母环形红孢子虫脂滴蛋白质组动力学。

Eukaryotic Cell Pub Date : 2015-03-01 Epub Date: 2015-01-09 DOI: 10.1128/EC.00141-14

Zhiwei Zhu, Yunfeng Ding, Zhiwei Gong, Li Yang, Sufang Zhang, Congyan Zhang, Xinping Lin, Hongwei Shen, Hanfa Zou, Zhensheng Xie, Fuquan Yang, Xudong Zhao, Pingsheng Liu, Zongbao K Zhao

{"title":"Dynamics of the lipid droplet proteome of the Oleaginous yeast rhodosporidium toruloides.","authors":"Zhiwei Zhu, Yunfeng Ding, Zhiwei Gong, Li Yang, Sufang Zhang, Congyan Zhang, Xinping Lin, Hongwei Shen, Hanfa Zou, Zhensheng Xie, Fuquan Yang, Xudong Zhao, Pingsheng Liu, Zongbao K Zhao","doi":"10.1128/EC.00141-14","DOIUrl":"https://doi.org/10.1128/EC.00141-14","url":null,"abstract":"Lipid droplets (LDs) are ubiquitous organelles that serve as a neutral lipid reservoir and a hub for lipid metabolism. Manipulating LD formation, evolution, and mobilization in oleaginous species may lead to the production of fatty acid-derived biofuels and chemicals. However, key factors regulating LD dynamics remain poorly characterized. Here we purified the LDs and identified LD-associated proteins from cells of the lipid-producing yeast Rhodosporidium toruloides cultured under nutrient-rich, nitrogen-limited, and phosphorus-limited conditions. The LD proteome consisted of 226 proteins, many of which are involved in lipid metabolism and LD formation and evolution. Further analysis of our previous comparative transcriptome and proteome data sets indicated that the transcription level of 85 genes and protein abundance of 77 proteins changed under nutrient-limited conditions. Such changes were highly relevant to lipid accumulation and partially confirmed by reverse transcription-quantitative PCR. We demonstrated that the major LD structure protein Ldp1 is an LD marker protein being upregulated in lipid-rich cells. When overexpressed in Saccharomyces cerevisiae, Ldp1 localized on the LD surface and facilitated giant LD formation, suggesting that Ldp1 plays an important role in controlling LD dynamics. Our results significantly advance the understanding of the molecular basis of lipid overproduction and storage in oleaginous yeasts and will be valuable for the development of superior lipid producers. ","PeriodicalId":11891,"journal":{"name":"Eukaryotic Cell","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2015-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1128/EC.00141-14","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32966769","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: 70

DNA double-strand breaks and telomeres play important roles in trypanosoma brucei antigenic variation. DNA双链断裂和端粒在布氏锥虫抗原变异中起重要作用。

Eukaryotic Cell Pub Date : 2015-03-01 Epub Date: 2015-01-09 DOI: 10.1128/EC.00207-14

Bibo Li

{"title":"DNA double-strand breaks and telomeres play important roles in trypanosoma brucei antigenic variation.","authors":"Bibo Li","doi":"10.1128/EC.00207-14","DOIUrl":"https://doi.org/10.1128/EC.00207-14","url":null,"abstract":"Human-infecting microbial pathogens all face a serious problem of elimination by the host immune response. Antigenic variation is an effective immune evasion mechanism where the pathogen regularly switches its major surface antigen. In many cases, the major surface antigen is encoded by genes from the same gene family, and its expression is strictly monoallelic. Among pathogens that undergo antigenic variation, Trypanosoma brucei (a kinetoplastid), which causes human African trypanosomiasis, Plasmodium falciparum (an apicomplexan), which causes malaria, Pneumocystis jirovecii (a fungus), which causes pneumonia, and Borrelia burgdorferi (a bacterium), which causes Lyme disease, also express their major surface antigens from loci next to the telomere. Except for Plasmodium, DNA recombination-mediated gene conversion is a major pathway for surface antigen switching in these pathogens. In the last decade, more sophisticated molecular and genetic tools have been developed in T. brucei, and our knowledge of functions of DNA recombination in antigenic variation has been greatly advanced. VSG is the major surface antigen in T. brucei. In subtelomeric VSG expression sites (ESs), VSG genes invariably are flanked by a long stretch of upstream 70-bp repeats. Recent studies have shown that DNA double-strand breaks (DSBs), particularly those in 70-bp repeats in the active ES, are a natural potent trigger for antigenic variation in T. brucei. In addition, telomere proteins can influence VSG switching by reducing the DSB amount at subtelomeric regions. These findings will be summarized and their implications will be discussed in this review. ","PeriodicalId":11891,"journal":{"name":"Eukaryotic Cell","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2015-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1128/EC.00207-14","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32966771","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: 40

The ADP/ATP carrier and its relationship to oxidative phosphorylation in ancestral protist trypanosoma brucei. 祖先原生布氏锥虫ADP/ATP载体及其与氧化磷酸化的关系。

Eukaryotic Cell Pub Date : 2015-03-01 Epub Date: 2015-01-23 DOI: 10.1128/EC.00238-14

Anna Gnipová, Karolína Šubrtová, Brian Panicucci, Anton Horváth, Julius Lukeš, Alena Zíková

{"title":"The ADP/ATP carrier and its relationship to oxidative phosphorylation in ancestral protist trypanosoma brucei.","authors":"Anna Gnipová, Karolína Šubrtová, Brian Panicucci, Anton Horváth, Julius Lukeš, Alena Zíková","doi":"10.1128/EC.00238-14","DOIUrl":"https://doi.org/10.1128/EC.00238-14","url":null,"abstract":"The highly conserved ADP/ATP carrier (AAC) is a key energetic link between the mitochondrial (mt) and cytosolic compartments of all aerobic eukaryotic cells, as it exchanges the ATP generated inside the organelle for the cytosolic ADP. Trypanosoma brucei, a parasitic protist of medical and veterinary importance, possesses a single functional AAC protein (TbAAC) that is related to the human and yeast ADP/ATP carriers. However, unlike previous studies performed with these model organisms, this study showed that TbAAC is most likely not a stable component of either the respiratory supercomplex III+IV or the ATP synthasome but rather functions as a physically separate entity in this highly diverged eukaryote. Therefore, TbAAC RNA interference (RNAi) ablation in the insect stage of T. brucei does not impair the activity or arrangement of the respiratory chain complexes. Nevertheless, RNAi silencing of TbAAC caused a severe growth defect that coincides with a significant reduction of mt ATP synthesis by both substrate and oxidative phosphorylation. Furthermore, TbAAC downregulation resulted in a decreased level of cytosolic ATP, a higher mt membrane potential, an elevated amount of reactive oxygen species, and a reduced consumption of oxygen in the mitochondria. Interestingly, while TbAAC has previously been demonstrated to serve as the sole ADP/ATP carrier for ADP influx into the mitochondria, our data suggest that a second carrier for ATP influx may be present and active in the T. brucei mitochondrion. Overall, this study provides more insight into the delicate balance of the functional relationship between TbAAC and the oxidative phosphorylation (OXPHOS) pathway in an early diverged eukaryote. ","PeriodicalId":11891,"journal":{"name":"Eukaryotic Cell","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2015-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1128/EC.00238-14","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32998929","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: 19

Functional complementation analyses reveal that the single PRAT family protein of trypanosoma brucei is a divergent homolog of Tim17 in saccharomyces cerevisiae. 功能互补分析表明，布鲁氏锥虫PRAT家族蛋白是酿酒酵母Tim17的分化同源物。

Eukaryotic Cell Pub Date : 2015-03-01 Epub Date: 2015-01-09 DOI: 10.1128/EC.00203-14

Ebony Weems, Ujjal K Singha, VaNae Hamilton, Joseph T Smith, Karin Waegemann, Dejana Mokranjac, Minu Chaudhuri

{"title":"Functional complementation analyses reveal that the single PRAT family protein of trypanosoma brucei is a divergent homolog of Tim17 in saccharomyces cerevisiae.","authors":"Ebony Weems, Ujjal K Singha, VaNae Hamilton, Joseph T Smith, Karin Waegemann, Dejana Mokranjac, Minu Chaudhuri","doi":"10.1128/EC.00203-14","DOIUrl":"https://doi.org/10.1128/EC.00203-14","url":null,"abstract":"Trypanosoma brucei, a parasitic protozoan that causes African trypanosomiasis, possesses a single member of the presequence and amino acid transporter (PRAT) protein family, which is referred to as TbTim17. In contrast, three homologous proteins, ScTim23, ScTim17, and ScTim22, are found in Saccharomyces cerevisiae and higher eukaryotes. Here, we show that TbTim17 cannot rescue Tim17, Tim23, or Tim22 mutants of S. cerevisiae. We expressed S. cerevisiae Tim23, Tim17, and Tim22 in T. brucei. These heterologous proteins were properly imported into mitochondria in the parasite. Further analysis revealed that although ScTim23 and ScTim17 were integrated into the mitochondrial inner membrane and assembled into a protein complex similar in size to TbTim17, only ScTim17 was stably associated with TbTim17. In contrast, ScTim22 existed as a protease-sensitive soluble protein in the T. brucei mitochondrion. In addition, the growth defect caused by TbTim17 knockdown in T. brucei was partially restored by the expression of ScTim17 but not by the expression of either ScTim23 or ScTim22, whereas the expression of TbTim17 fully complemented the growth defect caused by TbTim17 knockdown, as anticipated. Similar to the findings for cell growth, the defect in the import of mitochondrial proteins due to depletion of TbTim17 was in part restored by the expression of ScTim17 but was not complemented by the expression of either ScTim23 or ScTim22. Together, these results suggest that TbTim17 is divergent compared to ScTim23 but that its function is closer to that of ScTim17. In addition, ScTim22 could not be sorted properly in the T. brucei mitochondrion and thus failed to complement the function of TbTim17. ","PeriodicalId":11891,"journal":{"name":"Eukaryotic Cell","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2015-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1128/EC.00203-14","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32965267","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: 16

The krebs cycle enzyme α-ketoglutarate decarboxylase is an essential glycosomal protein in bloodstream African trypanosomes. 克雷布斯循环酶α-酮戊二酸脱羧酶是非洲锥虫血液中必需的糖体蛋白。

Eukaryotic Cell Pub Date : 2015-03-01 Epub Date: 2014-11-21 DOI: 10.1128/EC.00214-14

Steven Sykes, Anthony Szempruch, Stephen Hajduk

{"title":"The krebs cycle enzyme α-ketoglutarate decarboxylase is an essential glycosomal protein in bloodstream African trypanosomes.","authors":"Steven Sykes, Anthony Szempruch, Stephen Hajduk","doi":"10.1128/EC.00214-14","DOIUrl":"https://doi.org/10.1128/EC.00214-14","url":null,"abstract":"α-Ketoglutarate decarboxylase (α-KDE1) is a Krebs cycle enzyme found in the mitochondrion of the procyclic form (PF) of Trypanosoma brucei. The bloodstream form (BF) of T. brucei lacks a functional Krebs cycle and relies exclusively on glycolysis for ATP production. Despite the lack of a functional Krebs cycle, α-KDE1 was expressed in BF T. brucei and RNA interference knockdown of α-KDE1 mRNA resulted in rapid growth arrest and killing. Cell death was preceded by progressive swelling of the flagellar pocket as a consequence of recruitment of both flagellar and plasma membranes into the pocket. BF T. brucei expressing an epitope-tagged copy of α-KDE1 showed localization to glycosomes and not the mitochondrion. We used a cell line transfected with a reporter construct containing the N-terminal sequence of α-KDE1 fused to green fluorescent protein to examine the requirements for glycosome targeting. We found that the N-terminal 18 amino acids of α-KDE1 contain overlapping mitochondrion- and peroxisome-targeting sequences and are sufficient to direct localization to the glycosome in BF T. brucei. These results suggest that α-KDE1 has a novel moonlighting function outside the mitochondrion in BF T. brucei. ","PeriodicalId":11891,"journal":{"name":"Eukaryotic Cell","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2015-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1128/EC.00214-14","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32830126","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: 11

The selenocysteine tRNA gene in leishmania major is transcribed by both RNA polymerase II and RNA polymerase III. 利什曼原虫的硒代半胱氨酸tRNA基因由RNA聚合酶II和RNA聚合酶III转录。

Eukaryotic Cell Pub Date : 2015-03-01 Epub Date: 2014-12-29 DOI: 10.1128/EC.00239-14

Norma E Padilla-Mejía, Luis E Florencio-Martínez, Rodrigo Moreno-Campos, Juan C Vizuet-de-Rueda, Ana M Cevallos, Rosaura Hernández-Rivas, Rebeca Manning-Cela, Santiago Martínez-Calvillo

{"title":"The selenocysteine tRNA gene in leishmania major is transcribed by both RNA polymerase II and RNA polymerase III.","authors":"Norma E Padilla-Mejía, Luis E Florencio-Martínez, Rodrigo Moreno-Campos, Juan C Vizuet-de-Rueda, Ana M Cevallos, Rosaura Hernández-Rivas, Rebeca Manning-Cela, Santiago Martínez-Calvillo","doi":"10.1128/EC.00239-14","DOIUrl":"https://doi.org/10.1128/EC.00239-14","url":null,"abstract":"Eukaryotic tRNAs, transcribed by RNA polymerase III (Pol III), contain boxes A and B as internal promoter elements. One exception is the selenocysteine (Sec) tRNA (tRNA-Sec), whose transcription is directed by an internal box B and three extragenic sequences in vertebrates. Here we report on the transcriptional analysis of the tRNA-Sec gene in the protozoan parasite Leishmania major. This organism has unusual mechanisms of gene expression, including Pol II polycistronic transcription and maturation of mRNAs by trans splicing, a process that attaches a 39-nucleotide miniexon to the 5' end of all the mRNAs. In L. major, tRNA-Sec is encoded by a single gene inserted into a Pol II polycistronic unit, in contrast to most tRNAs, which are clustered at the boundaries of polycistronic units. 5' rapid amplification of cDNA ends and reverse transcription-PCR experiments showed that some tRNA-Sec transcripts contain the miniexon at the 5' end and a poly(A) tail at the 3' end, indicating that the tRNA-Sec gene is polycistronically transcribed by Pol II and processed by trans splicing and polyadenylation, as was recently reported for the tRNA-Sec genes in the related parasite Trypanosoma brucei. However, nuclear run-on assays with RNA polymerase inhibitors and with cells that were previously UV irradiated showed that the tRNA-Sec gene in L. major is also transcribed by Pol III. Thus, our results indicate that RNA polymerase specificity in Leishmania is not absolute in vivo, as has recently been found in other eukaryotes. ","PeriodicalId":11891,"journal":{"name":"Eukaryotic Cell","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2015-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1128/EC.00239-14","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"32940966","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: 9