Marek's disease virus (MDV) homologues of herpes simplex virus type 1 UL49 (VP22) and UL48 (VP16) genes: high-level expression and characterization of MDV-1 VP22 and VP16.
Genes UL49 and UL48 of Marek's disease virus 1 (MDV-1) strain RB1B, encoding the respective homologues of herpes simplex virus type 1 (HSV-1) genes VP22 and VP16, were cloned into a baculovirus vector. Seven anti-VP22 MAbs and one anti-VP16 MAb were generated and used to identify the tegument proteins in cells infected lytically with MDV-1. The two genes are known to be transcribed as a single bicistronic transcript, and the detection of only one of the two proteins (VP22) in MSB-1 lymphoma and in chicken embryo skin cells infected with MDV-1 prompted the study of the transcription/translation of the UL49-48 sequence in an in vivo and in vitro expression system. VP16 was expressed in vitro at detectable levels, whereas it could only be detected at a lower level in a more controlled environment. It was demonstrated that VP22 is phosphorylated in insect cells and possesses the remarkable property of being imported into all cells in a monolayer. VP22 localized rapidly and efficiently to nuclei, like its HSV-1 counterpart. The DNA-binding property of VP22 is also reported and a part of the region responsible for this activity was identified between aa 16 and 37 in the N-terminal region of the protein.
Dorange
Fabien
F
Laboratoire de Virologie Moléculaire, Station de Pathologie Aviaire et de Parasitologie, Centre INRA de Tours, 37380 Nouzilly, France1.
El Mehdaoui
Slimane
S
Laboratoire de Virologie Moléculaire, INSERM EMIV-00-10, Faculté des Sciences Pharmaceutiques 'Philippe Maupas', 37200 Tours, France2.
Pichon
Chantal
C
Centre de Biophysique Moléculaire, Glycobiologie, CNRS UPR4301 et Université d'Orleans, rue Charles-Sadron, F-45071 Orleans Cedex 02, France3.
Coursaget
Pierre
P
Laboratoire de Virologie Moléculaire, INSERM EMIV-00-10, Faculté des Sciences Pharmaceutiques 'Philippe Maupas', 37200 Tours, France2.
Vautherot
Jean-François
JF
Laboratoire de Virologie Moléculaire, Station de Pathologie Aviaire et de Parasitologie, Centre INRA de Tours, 37380 Nouzilly, France1.
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics*; D015967:Gene Expression Regulation, Viral; D018259:Herpesvirus 1, Human / Q000235:genetics*; D008381:Herpesvirus 2, Gallid / Q000235:genetics*; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D018411:Spodoptera; D013329:Structure-Activity Relationship; D014158:Transcription, Genetic; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics*; D015678:Viral Structural Proteins / Q000235:genetics*
D004279:DNA, Viral; C092965:UL48 protein, Gallid herpesvirus 2; D014764:Viral Proteins; D015678:Viral Structural Proteins; C073413:herpes simplex virus type 1 protein VP22; D004247:DNA