Ribosome and protein synthesis modifications after infection of human epidermoid carcinoma cells with herpes simplex virus type 1.
Modifications of ribosomes have been investigated in human epidermoid carcinoma-2 cells at different stages of herpes simplex virus type 1 infection. Very early in infection, there is an increase in ribosomal protein S6 phosphorylation even in the absence of serum. The same result is obtained in the presence of actinomycin D. At early infection time, ribosomal proteins S2, S3a and Sa are newly phosphorylated. At early and early-late times, three phosphorylated non-ribosomal proteins (v1, v2 and v3) are differently associated temporally to ribosomes. Analyses of proteins extracted from 40S subunits, 80S ribosomes and polysomes show that v1 and v2 are distributed differently among the different ribosomal populations. S6 phosphopeptides were found to be identical after serum stimulation and after viral infection. In every case phosphoserine and phosphothreonine were identified in S6. Only phosphoserine was found in other phosphorylated proteins. Our results indicate that herpes simplex virus type 1 is able to modify pre-existing ribosomes: (i) by stimulating a pre-existing kinase for S6 phosphorylation even in the absence of serum and of viral genome expression; (ii) by inducing new specific kinase activity(ies); and (iii) by association of new, phosphorylated proteins to ribosomes. These ribosomal modifications are correlated with changes in protein synthesis, as shown by two-dimensional electrophoretic analyses of newly synthesized 35S-labelled proteins.
Masse
T
T
Laboratoire de Biologie Moléculaire et Cellulaire, CNRS UMR 30, Faculté de Médecine Alexis Carrel, Lyon, France.
Garcin
D
D
Jacquemont
B
B
Madjar
J J
JJ
D002294:Carcinoma, Squamous Cell; D003470:Culture Media; D003609:Dactinomycin / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D010766:Phosphorylation; D014176:Protein Biosynthesis; D012269:Ribosomal Proteins / Q000032:analysis / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*; D018139:Simplexvirus / Q000502:physiology*; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000096:biosynthesis*
D003470:Culture Media; D012269:Ribosomal Proteins; D014764:Viral Proteins; D003609:Dactinomycin