RESUMEN
Substantial fraction of high-quality information is continuously being added into the existing pool of knowledge related to the biology of telomeres. Based on the insights gleaned from decades of research, it is clear that chromosomal stability needs a highly controlled and dynamic balance of DNA gain and loss in each terminal tract of telomeric repeats. Telomeres are formed by tandem repeats of TTAGGG sequences, which are gradually lost with each round of division of the cells. Targeted inhibition of telomerase to effectively induce apoptosis in cancer cells has attracted tremendous attention and overwhelmingly increasingly list of telomerase inhibitors truthfully advocates pharmacological significance of telomerase. Telomerase reverse transcriptase (TERT) is a multi-talented and catalytically active component of the telomerase-associated protein machinery. Different proteins of telomerase-associated machinery work in a synchronized and orchestrated manner to ensure proper maintenance of telomeric length of chromosomes. Rapidly emerging scientific findings about regulation of TERT by microRNAs has revolutionized our understanding related to the biology of telomeres and telomerase. In this review, we have comprehensively discussed how different miRNAs regulate TERT in different cancers. Use of miRNA-based therapeutics against TERT in different cancers needs detailed research in preclinical models for effective translation of laboratory findings to clinically effective therapeutics.
Asunto(s)
MicroARNs/metabolismo , Primula/enzimología , Primula/genética , Apoptosis/genética , Apoptosis/fisiología , MicroARNs/genética , Filogenia , Plastidios/genética , Telómero/genética , Telómero/metabolismoRESUMEN
Heterostyly is a wide-spread floral adaptation to promote outbreeding, yet its genetic basis and evolutionary origin remain poorly understood. In Primula (primroses), heterostyly is controlled by the S-locus supergene that determines the reciprocal arrangement of reproductive organs and incompatibility between the two morphs. However, the identities of the component genes remain unknown. Here, we identify the Primula CYP734A50 gene, encoding a putative brassinosteroid-degrading enzyme, as the G locus that determines the style-length dimorphism. CYP734A50 is only present on the short-styled S-morph haplotype, it is specifically expressed in S-morph styles, and its loss or inactivation leads to long styles. The gene arose by a duplication specific to the Primulaceae lineage and shows an accelerated rate of molecular evolution. Thus, our results provide a mechanistic explanation for the Primula style-length dimorphism and begin to shed light on the evolution of the S-locus as a prime model for a complex plant supergene.
Asunto(s)
Sistema Enzimático del Citocromo P-450/metabolismo , Flores/anatomía & histología , Proteínas de Plantas/metabolismo , Primula/anatomía & histología , Primula/enzimología , Sistema Enzimático del Citocromo P-450/genética , Evolución Molecular , Duplicación de Gen , Perfilación de la Expresión Génica , Silenciador del Gen , Primula/genética , PrimulaceaeRESUMEN
This manuscript describes an efficient analytical assay combining high-performance liquid chromatography with UV detection (HPLC-UV), liquid chromatography coupled with tandem mass spectrometry (LC-MS-MS), and gas chromatography with mass spectrometry (GC-MS) for the characterization and C=C bond localization on the long chain base of sphingolipids in yeast extracts in order to identify the plant sphingolipid desaturases activity. Samples of wild type control and transgenic yeast expressing putative sphingolipid desaturases were hydrolyzed into long chain bases. Mono-unsaturated long chain base, dehydrophytosphingosine (t18:1), in transgenic yeast as a result of the function of plant sphingolipid desaturase was detected with cis, trans-isomers resolution by reverse phase HPLC-UV as DNP (2,4-dinitrophenyl) derivatives along with saturated phytosphingosine (t18:0). The molecular structure of phytosphingosine was confirmed by negative-ion LC-MS-MS, which also served as a rapid tool for screening the plant spingolipid desaturase activity with 2-min run time under multiple-reaction monitoring (MRM) mode. The C=C bond location of dehydrophytosphingosine was further identified by GC-MS after being converted into picolinyl derivatives. This assay combines multiple chromatographic and mass spectrometric techniques with gentle chemical procedures to provide capacities for rapid determination of the plant sphingolipid desaturase activity as well as identification of their active sites in the backbone of the sphingolipid species in yeast.
Asunto(s)
Cromatografía Líquida de Alta Presión/métodos , Oxidorreductasas/química , Proteínas de Plantas/química , Primula/enzimología , Espectrometría de Masas en Tándem/métodos , Oxidorreductasas/genética , Oxidorreductasas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismoRESUMEN
cDNA clones encoding cytochrome b(5) fusion desaturases were isolated from Primula cortusoides L. and Primula luteola Ruprecht, species previously shown to preferentially accumulate either n-6 or n-3 Delta6-desaturated fatty acids, respectively. Functional characterisation of these desaturases in yeast revealed that the recombinant Primula enzymes displayed substrate preferences, resulting in the predominant synthesis of either gamma-linolenic acid (n-6) or stearidonic acid (n-3). Independent expression of the two Primula desaturases in transgenic Arabidopsis thaliana confirmed these results, with gamma-linolenic acid and stearidonic acid accumulating in both leaf and seed tissues to different levels, depending on the substrate specificity of the desaturase. Targeted lipid analysis of transgenic Arabidopsis lines revealed the presence of Delta6-desaturated fatty acids in the acyl-CoA pools of leaf but not seed tissue. The implications for the transgenic synthesis of C(20) polyunsaturated fatty acids via the elongation of Delta6-desaturated fatty acids are discussed, as is the potential of using Primula desaturases in the synthesis of C(18) n-3 polyunsaturated fatty acids such as stearidonic acid.
Asunto(s)
Ácido Graso Desaturasas/metabolismo , Primula/enzimología , Plantas Modificadas Genéticamente , Reacción en Cadena de la Polimerasa , Especificidad por SustratoRESUMEN
Fatty acid Delta(6)-desaturation, the first committed step in C(20) polyunsaturated fatty acid biosynthesis, is generally considered not to discriminate between n-3 and n-6 substrates. We previously identified higher plant species that showed preferential Delta(6)-desaturation of n-3 C(18) fatty acid substrates. A polymerase chain reaction-based approach was used to isolate 'front-end' cytochrome b(5) fusion desaturases from Primula vialii Franchet and Primula farinosa L. Functional analysis in Saccharomyces cerevisiae identified fatty acid Delta(6)-desaturases with a strong specificity for the n-3 substrate alpha-linolenic acid (18:3 Delta(9,12,15)). These results indicate that the accumulation of octadecatetraenoic acid (18:4 Delta(6,9,12,15)) in planta is due to the activity of a novel n-3-specific fatty acid Delta(6)-desaturase.