ABSTRACT
In order to identify host factors which interact with the movement protein (MP) of Apple chlorotic leaf spot virus (ACLSV), ACLSV MP was cloned into the bait vector pGBKT7 and used to screen a cDNA library of Malus sylvestris cv. R12740-7A, which had previously been constructed by yeast two-hybrid sequencing transformation. The protein functions of the identified host factors were determined according to their gene annotations in GenBank. The result showed that the bait plasmid pGBKT7-MP showed no virulence or self-activating effect on yeast strain Y2H Gold. Sixty-nine interactor proteins were identified, which were divided into the following 10 classes according to their described functions: hydrolases; pathogenesis-related proteins; DNA binding proteins; phosphatases; ligases; proteins with catalytic activity; phenylalanine ammonialyases; peroxidases; NAD binding proteins; and proteins of unknown function. Bioinformatic analysis of gene homology suggested that phosphatases, pathogenesis-related proteins and glyceraldehyde-3-phosphate dehydrogenase A may play an important role in the interaction between virus and host. This study may provide a theoretical basis for the further study of viral pathogenesis and virus-host interaction mechanisms.
Subject(s)
Flexiviridae , Genetics , Metabolism , Malus , Genetics , Metabolism , Virology , Molecular Sequence Data , Plant Diseases , Genetics , Virology , Plant Proteins , Genetics , Metabolism , Plant Viral Movement Proteins , Genetics , Metabolism , Protein Binding , Two-Hybrid System TechniquesABSTRACT
The Rad1 gene is evolutionarily conserved from yeast to human. The fission yeast Schizosaccharomyces pombe Rad1 ortholog promotes cell survival against DNA damage and is required for G(2)/M checkpoint activation. In this study, mouse embryonic stem (ES) cells with a targeted deletion of Mrad1, the mouse ortholog of this gene, were created to evaluate its function in mammalian cells. Mrad1 (-/-) ES cells were highly sensitive to ultraviolet-light (UV light), hydroxyurea (HU) and gamma rays, and were defective in G(2)/M as well as S/M checkpoints. These data indicate that Mrad1 is required for repairing DNA lesions induced by UV-light, HU and gamma rays, and for mediating G(2)/M and S/M checkpoint controls. We further demonstrated that Mrad1 plays an important role in homologous recombination repair (HRR) in ES cells, but a minor HRR role in differentiated mouse cells.
Subject(s)
Animals , Mice , Cell Division , Cell Proliferation , DNA Damage , DNA Repair , Embryonic Stem Cells , Metabolism , Exonucleases , Genetics , Metabolism , Physiology , G2 Phase , Gamma Rays , Gene Deletion , Hydroxyurea , Pharmacology , Ultraviolet RaysABSTRACT
Cold acclimation can improve freezing tolerance. Here cDNA amplified fragment length polymorphism (cDNA-AFLP) was used to isolate differentially expressed cDNAs and a Pp-LIM only A cDNA was isolated and identified in the cold acclimation of Physcomitrella patens. Real-time RT-PCR indicated it is obviously up-regulated at 6 h, 12 h, 24 h, 48 h and 72 h after cold acclimation. After comparing the cDNA with the gene sequence, seven introns and eight exons were identified in the cDNA. The cDNA putatively encodes a protein of 345 amino acid residues and only contains one LIM domain which has highly similarity with the PDZ/LIM domains of the protein family in animals. We proposed that Pp-LIM only A be a new gene coding for the LIM-domain containing protein and enhance stability of cell membrane via their effects on cytoskeleton during cold acclimation in P. patens.