Simian virus 40 T/t antigens and lamin A/C small interfering RNA rescue the phenotype of an Epstein-Barr virus protein kinase (BGLF4) mutant.
The Epstein-Barr virus (EBV)-encoded viral protein kinase, EBV-PK (the BGLF4 gene product), is required for efficient nuclear viral egress in 293 cells. However, since EBV-PK phosphorylates a number of different viral and cellular proteins (including lamin A/C), the relative importance of each target during lytic viral replication remains unclear. We show here that an EBV PK mutant (PKmut; containing stop codons at residues 1 and 5 in EBV-PK) is highly defective for release of infectious virus from 293 cells but not 293T cells. Furthermore, the phenotype of the PKmut in 293 cells is substantially reversed by expression of the simian virus 40 (SV40) large (T) and small (t) T antigens. Efficient rescue requires the presence of both SV40 T/t proteins. We show that 293T cells have a much higher level of constitutive lamin A/C phosphorylation than do 293 cells over residues (S22 and S392) that promote phosphorylation-dependent nuclear disassembly and that both large T and small t contribute to enhanced lamin A/C phosphorylation. Finally, we demonstrate that knockdown of lamin A/C expression using small interfering RNA also rescues the PKmut phenotype in 293 cells. These results suggest that essential roles of EBV-PK during lytic viral replication include the phosphorylation and dispersion of lamin A/C.
Meng
Qiao
Q
Department of Oncology, McArdle Laboratory for Cancer Research, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, WI 53706, USA.
Hagemeier
Stacy R
SR
Kuny
Chad V
CV
Kalejta
Robert F
RF
Kenney
Shannon C
SC
D000952:Antigens, Polyomavirus Transforming / Q000096:biosynthesis*; D002460:Cell Line; D055785:Gene Knockdown Techniques; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000254:growth & development*; D054884:Host-Pathogen Interactions; D006801:Humans; D034904:Lamin Type A / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000172:deficiency* / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics; D013539:Simian virus 40; D014764:Viral Proteins / Q000502:physiology*
D000952:Antigens, Polyomavirus Transforming; C494847:LMNA protein, human; D034904:Lamin Type A; D034741:RNA, Small Interfering; D014764:Viral Proteins; C406063:BGLF4 protein, Epstein-Barr virus; D017346:Protein Serine-Threonine Kinases