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1. UL28 and UL33 homologs of Marek’s disease virus terminase complex involved in the regulation of cleavage and packaging of viral DNA are indispensable for replication in cultured cells

Author

Luping Zheng, Yanan Wu, Gaiping Zhang, Yijie Xu, Rui Wang, Yongxiu Yao, Man Teng, Shuaikang Yang, Jun Luo, Guoqing Zhuang, Xiaojing Zhu, Aijun Sun, Venugopal Nair, Henan Agricultural University, and Henan Academy of Agricultural Sciences

Subjects
0301 basic medicine, animal structures, viruses, 030106 microbiology, Mutant, Chick Embryo, medicine.disease_cause, Virus Replication, Marek’s disease virus, Virus, 03 medical and health sciences, chemistry.chemical_compound, Viral Proteins, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], medicine, Animals, Amino Acid Sequence, Gene, UL28, RNA Cleavage, Marek's disease, lcsh:Veterinary medicine, Endodeoxyribonucleases, [SDV.BA.MVSA]Life Sciences [q-bio]/Animal biology/Veterinary medicine and animal Health, General Veterinary, biology, DNA packaging, Transfection, biology.organism_classification, Molecular biology, 3. Good health, Specific Pathogen-Free Organisms, Co-transfection, Mardivirus, 030104 developmental biology, Herpes simplex virus, chemistry, Viral replication, DNA, Viral, embryonic structures, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, lcsh:SF600-1100, Receptors, Chemokine, UL33, Sequence Alignment, DNA, Research Article
Abstract

Processing and packaging of herpesvirus genomic DNA is regulated by a packaging-associated terminase complex comprising of viral proteins pUL15, pUL28 and pUL33. Marek’s disease virus (MDV) homologs UL28 and UL33 showed conserved functional features with high sequence identity with the corresponding Herpes simplex virus 1 (HSV-1) homologs. As part of the investigations into the role of the UL28 and UL33 homologs of oncogenic MDV for DNA packaging and replication in cultured cells, we generated MDV mutant clones deficient in UL28 or UL33 of full-length MDV genomes. Transfection of UL28- or UL33-deleted BAC DNA into chicken embryo fibroblast (CEF) did not result either in the production of visible virus plaques, or detectable single cell infection after passaging onto fresh CEF cells. However, typical MDV plaques were detectable in CEF transfected with the DNA of revertant mutants where the deleted genes were precisely reinserted. Moreover, the replication defect of the UL28-deficient mutant was completely restored when fragment encoding the fullUL28gene was co-transfected into CEF cells. Viruses recovered from the revertant construct, as well as by theUL28co-transfection, showed replication ability comparable with parental virus. Furthermore, the transmission electron microscopy study indicated that immature capsids were assembled without the UL28 expression, but with the loss of infectivity. Importantly, predicted three-dimensional structures of UL28 between MDV and HSV-1 suggests conserved function in virus replication. For the first time, these results revealed that both UL28 and UL33 are essential for MDV replication through regulating DNA cleavage and packaging.

Published
2021

2. Total synthesis of a functional designer eukaryotic chromosome

Author

Eric M. Cooper, Peter Deng, Karen I. Zeller, Zheng Kuang, Aaron M. Moore, Jessica S. Dymond, Jef D. Boeke, Sindurathy Murugan, Judy Doong, Jaime Liu, Jeffrey S. Han, Kristie Charoen, Neta Agmon, Apurva Yeluru, Leslie A. Mitchell, Lisa Z. Scheifele, Jonathan P. Ling, Zheyuan Guo, Henry Ma, Marina Paul, Kimberly M. Cirelli, Romain Koszul, Charlotte E. Floria, Isabel E. Ishizuka, Joy Chang, Mariya London, Matthew G. Rubashkin, Sarah M. Richardson, Wei Xie, Jennifer Tullman, Christopher Fernandez, Judy Qiu, Kristin M. Boulier, Alex Rhee, Allison Suarez, Pablo A. Lee, Ruchi Patel, Remus S. Wong, Jonathan Liu, Joel S. Bader, Pavlo Bohutski, J. Andrew Martin, Brian J. Capaldo, Giovanni Stracquadanio, Venkatesh Srinivas, Sean Li, Jessica Mao, Allen T. Yu, Woo Jin Choi, Ina Y. Soh, Javaneh Jabbari, Katrina Caravelli, Yijie Xu, Jason I. Feinberg, Gilles Fischer, Kun Yang, Karthikeyan Kandavelou, Yu Ouyang, Nathaniel E. Sotuyo, Murat Bilgel, Matthias E. Linder, Narayana Annaluru, Andrew D. Wong, Jessilyn Dunn, Pasha Hadidi, James E. DiCarlo, Won Chan Oh, Jessica E. McDade, David Gladowski, Héloïse Muller, Denise Lin, Srinivasan Chandrasegaran, Alexandra McMillan, Sivaprakash Ramalingam, Calvin Y. L. Lau, Viktoriya London, Laura C. Paulsen, Nicolas Agier, Yizhi Cai, Michalis Hadjithomas, Department of Environmental Health Sciences [JHU Baltimore], Johns Hopkins University (JHU) School of Public Health, Régulation spatiale des Génomes - Spatial Regulation of Genomes, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), High Throughput Biology Center [Baltimore], Johns Hopkins University School of Medicine [Baltimore], New York University Langone Medical Center, Johns Hopkins University ( JHU ), Department of Biomedical Engineering and Institute of Genetic Medicine, Whiting School of Engineering, Biological Sciences, Research and Exploratory Development Department [Laurel], Johns Hopkins University Applied Physics Laboratory [Laurel, MD] ( APL ), Department of Biology [Loloya U. Baltimore], Loyola University Maryland, University of Edinburgh, Carnegie Institution for Science [Washington], Department of Biology [JHU Baltimore], Krieger School of Arts and Sciences [Baltimore], Whiting School of Engineering [Baltimore], Pondicherry Biotech Private Limited, Génomique des Microorganismes ( LGM ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Centre National de la Recherche Scientifique ( CNRS ), This work was supported by grants from NSF (MCB 0718846) to J.D.B., J.S.B., and S.C. and from Microsoft to J.S.B. S.M. and S.C were supported by a grant from NIH (GM077291 to S.C.), H. Muller, by a fellowship from Fondation pour la Recherche Médicale and a Pasteur-Roux fellowship, S.R., by an Exploratory Research Grant from the Maryland Stem Cell Research Fund, L.A.M., by a fellowship from the National Sciences and Engineering Research Council of Canada, S.M.R., by a fellowship from the U.S. Department of Energy, and J.S.D., by a fellowship from JHU Applied Physics Laboratory., Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), New York University Langone Medical Center (NYU Langone Medical Center), NYU System (NYU), Johns Hopkins University (JHU), Johns Hopkins University Applied Physics Laboratory [Laurel, MD] (APL), Loyola University [Maryland, Baltimore], Génomique des Microorganismes (LGM), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), and Carnegie Institution for Science

Subjects
Transposable element, [SDV]Life Sciences [q-bio], Genes, Fungal, Molecular Sequence Data, Saccharomyces cerevisiae, Bacterial genome size, [ SDV.BBM.BM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Biology, medicine.disease_cause, Polymerase Chain Reaction, Genome, Genomic Instability, Article, 03 medical and health sciences, Transformation, Genetic, 0302 clinical medicine, RNA, Transfer, medicine, DNA, Fungal, Gene, Sequence Deletion, 030304 developmental biology, Genetics, 0303 health sciences, Mutation, Multidisciplinary, Base Sequence, [ SDV ] Life Sciences [q-bio], Chromosome, RNA, Fungal, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Sequence Analysis, DNA, biology.organism_classification, Introns, Eukaryotic chromosome fine structure, Synthetic Biology, Genetic Fitness, Genome, Fungal, Chromosomes, Fungal, 030217 neurology & neurosurgery
Abstract

Designer Chromosome One of the ultimate aims of synthetic biology is to build designer organisms from the ground up. Rapid advances in DNA synthesis has allowed the assembly of complete bacterial genomes. Eukaryotic organisms, with their generally much larger and more complex genomes, present an additional challenge to synthetic biologists. Annaluru et al. (p. 55 , published online 27 March) designed a synthetic eukaryotic chromosome based on yeast chromosome III. The designer chromosome, shorn of destabilizing transfer RNA genes and transposons, is ∼14% smaller than its wild-type template and is fully functional with every gene tagged for easy removal.

Published
2014

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