Juan Manuel González-Prieto

The UmGcn5 gene encoding histone acetyltransferase from Ustilago maydis is involved in dimorphism and virulence

We isolated a gene encoding a histone acetyltransferase from Ustilago maydis (DC.) Cda., which is orthologous to the Saccharomyces cerevisiae GCN5 gene. The gene was isolated from genomic clones identified by their specific hybridization to a gene fragment obtained by the polymerase chain reaction (PCR). This gene (Umgcn5; um05168) contains an open reading frame (ORF) of 1421bp that encodes a putative protein of 473 amino acids with a Mr. of 52.6kDa. The protein exhibits a high degree of homology with histone acetyltransferases from different organisms. Null a2b2 ΔUmgcn5 mutants were constructed by substitution of the region encoding the catalytic site with a hygromycin B resistance cassette. Null a1b1 ΔUmgcn5 mutants were isolated from genetic crosses of a2b2 ΔUmgcn5 and a1b1 wild-type strains in maize. Mutants displayed a slight reduction in growth rate under different conditions, and were more sensitive than the wild type to stress conditions, but more important, they grew as long mycelial cells, and formed fuzz-like colonies under all conditions where wild-type strains grew in the yeast-like morphology and formed smooth colonies. This phenotype was not reverted by cAMP addition. Mutants were not virulent to maize plants, and were unable to form teliospores. These phenotypic alterations of the mutants were reverted by their transformation with the wild-type gene.

Transcriptomic analysis of the GCN5 gene reveals mechanisms of the epigenetic regulation of virulence and morphogenesis in Ustilago maydis

Chromatin in the eukaryotic nucleus is highly organized in the form of nucleosomes where histones wrap DNA. This structure may be altered by some chemical modifications of histones, one of them, acetylation by histone acetyltransferases (HATs) that originates relaxation of the nucleosome structure, providing access to different transcription factors and other effectors. In this way, HATs regulate cellular processes including DNA replication, and gene transcription. Previously, we isolated Ustilago maydis mutants deficient in the GCN5 HAT that are avirulent, and grow constitutively as mycelium. In this work, we proceeded to identify the genes differentially regulated by GCN5, comparing the transcriptomes of the mutant and the wild type using microarrays, to analyse the epigenetic control of virulence and morphogenesis. We identified 1203 genes, 574 positively and 629 negatively regulated in the wild type. We found that genes belonging to different categories involved in pathogenesis were downregulated in the mutant, and that genes involved in mycelial growth were negatively regulated in the wild type, offering a working hypothesis on the epigenetic control of virulence and morphogenesis of U. maydis. Interestingly, several differentially regulated genes appeared in clusters, suggesting a common regulation. Some of these belonged to pathogenesis or secondary metabolism.

Isolation and molecular analysis of Umhda2 a gene encoding a histone deacetylase from Ustilago maydis.

By use of the polymerase chain reaction and synthetic oligonucleotides designed from conserved regions, we amplified a fragment of a gene from Ustilago maydis encoding a putative histone deacetylase. With this probe we isolated the full gene from a minigenomic library. The gene (designated as Umhda2) contains an open reading frame (ORF) of 1701 bp encoding a protein of 566 amino acids. Multiple comparison analysis with other histone deacetylases suggests that the Umhda2 gene product belongs to the Rpd3-related family of proteins. The highest degree of homology with histone deacetylases from other organisms corresponded to Hda1p of Schizosaccharomyces pombe and Rpd3p of Saccharomyces cerevisiae with 64.2 and 62.2% of sequence similarity, respectively. It displayed a substantially lower similarity with another histone deacetylase from U. maydis (Hda1p, 52.4%). Semi-quantitative RTPCR results indicate that the gene is transcriptionally up-regulated during the in  vitro yeast-to-mycelium dimorphic transition.

Epigenetics: from the past to the present

ABSTRACT The definition of epigenetics is still under intense debate; however, its concept has evolved since it was originally introduced in 1939 by Conrad Hal Waddington as a way to reconcile antagonistic views between the school of preformationism and the school of epigenesis. The characterization of a large number of phenomena that diverge from the dogmas of classical genetics, and the discovery of the molecular mechanisms through which these phenomena occur, has given rise to a new area of study with important implications for biological sciences. Interactions between the environment and the DNA through modifications on the chromatin are not only responsible for the expression of a normal phenotype, these may be involved in the development of various pathologies. The epigenome, as the bridge between the genome and the phenotype, is no doubt one of the most interesting current ideas in genetics and is so revolutionary that it may change our present notions about inheritance and evolution. In this review, we made a compilation of the most important events in the history of epigenetics, its implications and some perspectives to the future. Abbreviations: DNA: deoxyribonucleic acid; RNA: ribonucleic acid; DNMT: DNA methyltransferase; MBP: methyl-CpG-binding proteins; HAT: histone acetyltransferase; HDAC: histone deacetylase; SAM: S-adenosyl methionine; ncRNA: non-coding RNA; rRNA: ribosomal RNA; miRNA: microRNA; siRNA: small interfering RNA; piRNA: Piwi-interacting RNA; XiRNA: X-inactivation RNA; lncRNA: long non-coding RNA; GR: glucocorticoid receptor; IGF2: insulin-like growth factor II; HPA: hypothalamic–pituitary–adrenal; TSA: trichostatin A; LINE: long interspersed nuclear elements; LOI: loss of genomic imprinting; MAS: McCune–Albright syndrome; AS: Angelman syndrome; PWS: Prader–Willi syndrome; FDA: Food and Drug Administration; AHEAD: International Human Epigenome Project; HEP: Human Epigenome Project; TMG: thiomethyl-β-D-galactoside

Climatic adaptation and ecological descriptors of wild beans from Mexico

Abstract Despite its economic, social, biological, and cultural importance, wild forms of the genus Phaseolus are not well represented in germplasm banks, and they are at great risk due to changes in land use as well as climate change. To improve our understanding of the potential geographical distribution of wild beans (Phaseolus spp.) from Mexico and support in situ and ex situ conservation programs, we determined the climatic adaptation ranges of 29 species and two subspecies of Phaseolus collected throughout Mexico. Based on five biotic and 117 abiotic variables obtained from different databases—WorldClim, Global‐Aridity, and Global‐PET—we performed principal component and cluster analyses. Germplasm was distributed among 12 climatic types from a possible 28. The general climatic ranges were as follows: 8–3,083 m above sea level; 12.07–26.96°C annual mean temperature; 10.33–202.68 mm annual precipitation; 9.33–16.56 W/m2 of net radiation; 11.68–14.23 hr photoperiod; 0.06–1.57 aridity index; and 10–1,728 mm/month of annual potential evapotranspiration. Most descriptive variables (25) clustered species into two groups: One included germplasm from semihot climates, and the other included germplasm from temperate climates. Species clustering showed 45% to 54% coincidence with species previously grouped using molecular data. The species P. filiformis, P. purpusii, and P. maculatus were found at low‐humidity locations; these species could be used to improve our understanding of the extreme aridity adaptation mechanisms used by wild beans to avoid or tolerate climate change as well as to introgress favorable alleles into new cultivars adapted to hot, dry environments.

Population Structure of Digestive Trypsin Phenotypes in Hatcheries for Pacific White Shrimp and Their Frequencies during Growth in Commercial Culture

AbstractTrypsin from Pacific white shrimp Litopenaeus vannamei is a polymorphic protein composed of three alleles (A, B, and C). The C allele is monomorphic for the first region, while A and B are allelic variations for the second region that are segregated according to Mendelian rules. These three alleles give rise to three individual trypsin phenotypes displayed in sodium dodecyl sulfate polyacrylamide gel electrophoresis: CBA, CB, and CA. The expected frequencies from mating CBA males and females resulted in offspring that consisted of 25% CB, 25% CA, and 50% CBA trypsin phenotypes. To define a management protocol for studying shrimp performance associated with the trypsin phenotype, the goal of this study was to determine if the expected proportions of the three trypsin phenotypes are represented in the population structure at breeding age in 11 hatcheries. Here, we hypothesized that the population structure of the trypsin phenotypes is represented by frequencies that are subject to selection, and it ...