Geminiviral protein Rep interferes in PCNA sumoylation

Rep is a multifunctional protein essential for replication of geminivirus that interferes with the sumoylation of a key protein in the DNA replication, PCNA (Proliferating Cell Nuclear Antigen). It is known that Rep is capable of interacting with a plethora of plant proteins, including PCNA. Despite the biological significance remains unknown, it’s thought that this interaction should play a key role for generating new copies of the virus genome. Therefore, in order to characterize this interaction, we study which lysines are sumoylated in tomato PCNA (SlPCNA). Considering conservation, location and presence of sumoylation domain criteria, we have identified some candidate lysines and studied how its mutation affects this protein sumoylation in Escherichia coli assays. Finally, we plan to confirm and characterize the Rep interference on SlPCNA sumoylation and determine if this interference occurs in planta.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

La proteína Rep/AL1 de geminivirus altera la sumoilación de PCNA

Los geminivirus, llamados así por la forma icosaédrica de su cápside, son una familia de virus patógenos de plantas que causan algunas de las enfermedades con mayor impacto económico a nivel mundial. Estos pequeños virus de ADN se replican en el núcleo de las células vegetales utilizando la maquinaria celular del hospedador, además de requerir la presencia de la proteína viral Rep. Rep es la única proteína del genoma del virus imprescindible para su replicación y es capaz de interaccionar con una gran variedad de proteínas del huésped. Trabajos previos de nuestro grupo demostraron que dos de esas proteínas son PCNA (Proliferating cell nuclear antigen), esencial en el metabolismo del DNA, y SCE (SUMO Conjugating Enzime), enzima que interviene en la sumoilación, uno de los principales mecanismos de modificación postraduccional que está implicado en la respuesta de la planta a estrés abiótico, en el desarrollo, el crecimiento y la respuesta a patógenos. Además, otros resultados que hemos obtenido demuestran que la expresión en Escherichia coli de la proteína Rep del begomovirus TGMV (Tomato golden mosaic virus) reduce la sumoilación de PCNA en plantas y que dicha interferencia no depende de la interacción de la proteína viral con SCE1. Para profundizar en el conocimiento de la interacción Rep-PCNA nos hemos propuesto identificar las lisinas de PCNA que se sumoilan. Teniendo en cuenta criterios de conservación, localización y presencia de dominio de sumoilación se han identificado varias lisinas candidatas y se ha estudiado cómo su mutación afecta a la sumoilación de la proteína. También nos hemos planteado confirmar y caracterizar la interferencia de Rep sobre la sumoilación de PCNA y determinar si dicha interferencia se produce en la planta.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Geminivirus replication protein decreases PCNA sumoylation at two acceptor sites

Geminiviruses are plant viruses with circular, single-stranded DNA (ssDNA) genomes that infect a broad range of plants causing substantial crop diseases worldwide. They replicate in nuclei of infected cells by using host DNA replication machinery and an essential protein encoded in their genome designated Rep (replication-associated protein). This multifunctional protein induces the accumulation of the host factors involved in replication and it is capable of interacting with a lot of plant proteins including PCNA (Proliferating Cell Nuclear Antigen), a processivity factor that coordinates a wide range of processes involved in maintenance, duplication and transmission of the genome, and the sumoylation enzyme that conjugates SUMO to target proteins (SUMO-conjugating enzyme- SCE). PCNA modification by SUMO, and also ubiquitin, has long been known to be of key importance for determining how DNA damage is processed by the replisome and for maintenance of overall genome integrity. In yeast, PCNA sumoylation has been associated to DNA repair involving homologous recombination (HR). Previously, we reported that Rep ectopic expression does not result in broad changes in the sumoylation pattern of plant cells, but it modifies the sumoylation state of selected host proteins. In this work, we show, using a reconstituted sumoylation system in Escherichia coli, that tomato PCNA is sumoylated at two residues, K254 and K164, and that co-expression of the Rep protein suppresses PCNA sumoylation at these lysines. Finally, we confirm that PCNA is sumoylated and that Rep also interferes with PCNA sumoylation in planta.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec

Analysis of Game Actions and Performance in Young Soccer Players: A Study Using Sequential Analysis

The objective of this research is to analyze the performance of actions in a reduced game situation in a sample of young soccer players. This is a game format in which possession of the ball has to be maintained. The sample consisted of 85 young players aged between 12 and 16 years, observing a total of 58 reduced games and using an analysis instrument called the Game Performance Assessment Instrument (GPAI). The essential data quality analyses are carried out, including the use of correlation indexes, Cohen’s Kappa and the Phi index for intra- and inter-observer reliability. Generalizability and invariance analyses are also performed to estimate the reliability, validity and precision of the numerical structure and its generalizability to other samples or populations with similar distributions. An observational design of nomothetic, punctual and multidimensional types is used, and subsequently, a sequential analysis of the game actions is carried out from the observations. The results show significant relationships between decision-making behaviors (adequate and inadequate) and technical execution behaviors (adequate and inadequate). The findings have highlighted a clear relationship of interdependence between technical execution and decision making. This information is very useful for the design and planning of training oriented to the optimization of sport performance in soccer.Partial funding for open access charge: Universidad de Málag

Cognitive functioning, physical fitness, and game performance in a sample of adolescent soccer players.

The aim of this study was to analyze the relationships between cognitive functioning, physical fitness, and game performance in a sample of adolescent soccer players. Eighty-five boys from a soccer team in Rincon de la Victoria (Malaga, Spain), aged between 12 and 16 years, participated in the study. The D2 and Global-Local Attention (GLA) tests were used to evaluate selective and divided attention, respectively, and the Coding and Symbol Search tests of the Wechsler Intelligence Scale for Children (WISC-IV) were used for processing speed. To analyze physical fitness, the standing long jump test, speed test, and Course-Navette test of the Alpha-Fitness battery were used. Finally, the Game Performance Assessment Instrument (GPAI) was used to analyze sports performance in three vs. three small-sided games. The analyses showed that cognitive functioning was related to game performance. The results obtained suggest the importance of cognitive functions for performance in this sport