Infectious disease research and one health education programs

One Health refers to the interconnected nature of the health and well-being of people, animals and the environments that they share. Central to this idea, is the realization that adverse health-effects in one area are linked to adverse effects in other area. Therefore, sustainable human health is intimately linked with sustainable animal and environmental health. This panel will include discussions of important and emerging aspects of One Health by experts in this field, each with specializations in different, but complementary, aspects of One Health that covers a wide range of disciplines and areas of expertise

The rem Mutations in the ATP-Binding Groove of the Rad3/XPD Helicase Lead to Xeroderma pigmentosum-Cockayne Syndrome-Like Phenotypes

The eukaryotic TFIIH complex is involved in Nucleotide Excision Repair and transcription initiation. We analyzed three yeast mutations of the Rad3/XPD helicase of TFIIH known as rem (recombination and mutation phenotypes). We found that, in these mutants, incomplete NER reactions lead to replication fork breaking and the subsequent engagement of the homologous recombination machinery to restore them. Nevertheless, the penetrance varies among mutants, giving rise to a phenotype gradient. Interestingly, the mutations analyzed reside at the ATP-binding groove of Rad3 and in vivo experiments reveal a gain of DNA affinity upon damage of the mutant Rad3 proteins. Since mutations at the ATP-binding groove of XPD in humans are present in the Xeroderma pigmentosum-Cockayne Syndrome (XP-CS), we recreated rem mutations in human cells, and found that these are XP-CS-like. We propose that the balance between the loss of helicase activity and the gain of DNA affinity controls the capacity of TFIIH to open DNA during NER, and its persistence at both DNA lesions and promoters. This conditions NER efficiency and transcription resumption after damage, which in human cells would explain the XP-CS phenotype, opening new perspectives to understand the molecular basis of the role of XPD in human disease.Research was funded by grants from the Spanish Ministry of Economy and Competitiveness (BFU2010-16372), the Junta de Andalucía (CVI4567) and the European Union (FEDER).Peer reviewe

Analysis of TFIIH recruitment to promoters in <i>rad3</i> mutants.

<p>(<b>A</b>) Chromatin Immunoprecipitation (ChIP) analysis of Tfb4-TAP. Cells were grown in synthetic complete (SC) medium until the exponential phase. ChIP analysis was performed at the <i>ALG9</i> promoter and normalized with respect to the <i>MFA2</i> promoter in <i>MAT</i><b>α</b> cells, which is constitutively repressed. (<b>B</b>) ChIP analysis of Tfb4-TAP after UV damage. Cells were grown in SC medium until the exponential phase, and then irradiated with 80 J/m². Analysis of the different time-point samples was performed at the <i>GRX1</i> promoter and normalized with respect to the <i>MFA2</i> promoter in <i>MAT</i><b>α</b> cells, which is constitutively repressed. The mean and the SD of triplicate assays of four independent experiments are depicted for each condition. *, p<0.05, **, p<0.01 (Student's t-test).</p

Hyper-recombination and genetic interactions of <i>rem</i> mutations with recombination and replication functions.

<p>(<b>A</b>) Analysis of Rad52 foci after exposure to UV-C. Only cells in S and G2 were considered. Error bars represent the SD of three independent experiments. One representative experiment is shown for <i>rad3-102</i> in order to facilitate comparison with previous published data <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1004859#pgen.1004859-MorielCarretero1" target="_blank">[15]</a>. (<b>B</b>) Analysis of genetic interactions of <i>rem</i> alleles of <i>RAD3</i> with <i>mre11Δ</i>, <i>rad51Δ</i> and <i>pol32Δ</i>. Tetrads dissected on rich medium are shown. The sites of double and triple mutants are indicated by triangles and squares, respectively. (<b>C</b>) Survival curves of WT, <i>rad3-101</i>, <i>rad51Δ</i> and their corresponding double mutant cells after UV-C exposure. (<b>D</b>) FACS profiles of mid-log cultures from WT, <i>rad3-101</i>, <i>rad51Δ</i> and their corresponding double mutant cells taken every 2 h after addition of 40 mM HU.</p

<i>rad3</i> mutants display a gradient response to UV irradiation.

<p>(<b>A</b>) Survival curves of WT and different yeast <i>rad3</i> mutants after UV-C exposure. (<b>B</b>) FACS profiles from WT, <i>rad3-101</i>, <i>rad3-107</i> and <i>rad3-102</i> cells synchronized in G1 with α-factor, untreated or UV-irradiated with 40 or 100 J/m² and released after 2 h. (<b>C</b>) Pulsed-field gel electrophoresis (PFGE) of DNA from WT, <i>rad3-101</i>, <i>rad3-107</i> and <i>rad3-102</i> cells synchronized in G1 with α-factor and further released into S phase. Bands reveal chromosome VII by hybridization with a probe of the <i>ADE5,7 locus</i>. Nonlinear (NLC) and full-length linear (FLC) chromosomes include replication intermediates, in the well, and pre- and post-replicated chromosomes, which enter the gel, respectively. Bars represent the quantification of NLC with respect to the total of signal of each lane. The bottom part of the membrane below the FLC is not shown since no signals, as expected from broken DNA molecules, were revealed by hybridization in any lane. (<b>D</b>) All details as in (C) except that after G1 synchronization, cells were UV-irradiated with 40 J/m² or 100 J/m² and released into S phase 2 h later.</p