Skip to main page content
U.S. flag

An official website of the United States government

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1998 Jun;112(3):482-9.
doi: 10.1046/j.1365-2249.1998.00605.x.

The human autoantigen La/SS-B accelerates herpes simplex virus type 1 replication in transfected mouse 3T3 cells

M Bachmann  1 , H Deister, A Pautz, J Laubinger, M Schmitz, D Falke, J Podlech, D Grölz
Affiliations

The human autoantigen La/SS-B accelerates herpes simplex virus type 1 replication in transfected mouse 3T3 cells

M Bachmann et al. Clin Exp Immunol. 1998 Jun.

Abstract

Permanently transfected mouse cell lines which expressed different levels of the human autoantigen La/SS-B were infected with different strains of herpes simplex virus type 1, including the strains ANG, HSZP, 17syn+ and HFEM. During infection the localization of the human La protein was followed using an anti-La MoAb, which recognized only the human La protein but did not cross-react with either the endogenous mouse La protein or any viral encoded protein. After infection La protein was transported from the nucleus to the cytoplasm. The time course of translocation was dependent on the amount of human La protein expressed in the respective cell line. Moreover, acceleration of viral replication was dependent on the level of expression of human La protein, suggesting that La protein is a cellular factor that facilitates virus replication.

PubMed Disclaimer

Figures

Fig. 3
Fig. 3
SDS–PAGE/immunoblotting analysis of mouse cell lines expressing the human La gene or either exon 1 or exon 1′ La mRNAs. Total extracts of mouse cells were analysed. Untransfected mouse 3T3 cells (3T3, lane a); cell lines expressing the human La gene (lane b), or human exon 1 La mRNA (lane c), or human exon 1′ La mRNA (lane d).
Fig. 1
Fig. 1
Epifluorescence analysis of mouse 3T3 cells permanently expressing human exon 1 (a,c,e,g) or 1′ La mRNAs (b,d,f,h). The cells were infected with herpes simplex virus type 1 strain ANG. (i,j) Untransfected mouse 3T3 cells. The cells were obtained 1 h post-infection (p.i.) (a,b,i) or 4 h p.i. (c,d,j), or 8 h p.i. (e,f) or 24 h p.i. (g,h).
Fig. 4
Fig. 4
Epifluorescence analysis of mouse 3T3 cells permanently expressing human exon 1′ (a,c,e) or 1′ La mRNAs (b,d,f). The cells were infected with herpes simplex virus type 1 strain HFEM (a,b), strain HSZP (c,d) or strain 17syn+ (e,f). The cells were obtained 8 h p.i. (a,b,e,f) or 6 h p.i. (c,d).
Fig. 2
Fig. 2
SDS–PAGE/immunoblotting analysis of mouse cells expressing the human exon 1 La mRNAs after infection with herpes simplex type 1. Total extracts of mouse cells were analysed. Untransfected mouse 3T3 cells (3T3, lanes a, c, f, i); cell lines expressing the exon 1 La mRNA under control of the cytomegalovirus (CMV) promoter (lanes b, d, e, g, h, j, k). The cells were infected with the herpes simplex virus type 1 strain ANG ((+), lanes c, e, f, h, i, k) or not infected ((–), lanes d, g, j). Cells were analysed 1 h (lanes c, d, e), 4 h (lanes f, g, h) or 8 h (lanes i, j, k) p.i. M, Marker, lane 1 = 97.4 kD, 2 = 66 kD, 3 = 43 kD, 4 = 31 kD, 5 = 21.5 kD, 6 = 14.5 kD.
Fig. 5
Fig. 5
Phase contrast analysis of mouse 3T3 cells after herpes simplex type 1 virus infection. The cells were infected with herpes simplex virus type 1 strain ANG. Untransfected mouse 3T3 cells (a), or cells expressing the exon 1′ under cytomegalovirus (CMV) promoter control (b), or cells expressing the human La gene under control of the genuine promoter elements (d), or cells expressing the exon 1 La mRNA under CMV promoter control (c) were fixed 5 h p.i. and phase contrast images were taken.
Fig. 6
Fig. 6
Epifluorescence analysis of cells stained with a MoAb to a surface protein of herpes simplex virus type 1. The cells were infected with herpes simplex virus strain ANG. Six hours p.i. the cells were stained with the MoAb HSV1-06 (see Materials and Methods). (a) Untransfected mouse 3T3 cells. (b) Cells transfected with the exon 1′ construct. (c) Cells transfected with the exon 1 construct. (d) Cells transfected with the human La gene.
Fig. 7
Fig. 7
Estimation of herpes virus DNA polymerase activity. Viral DNA polymerase activity was measured in transfected cell lines 2 h, 4 h, 6 h and 8 h post-infection (p.i.). Viral DNA polymerase activity was estimated using a commercially available ELISA system (see Materials and Methods). b, hatched bar, activity of Klenow enzyme in the presence of ammonium sulphate; ▪, neomycin-resistant cell line; hatched bars, exon 1′ La mRNA-expressing cell line; □, cell line expressing the human La gene. OD, Optical density. Error bars of two assays are given.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Tan EM. Antinuclear antibodies: diagnostic markers for autoimmune diseases and probes for cell biology. Adv Immunol. 1989;44:93–151. - PubMed
    1. Hendrick JP, Wolin S, Rinke J, Lerner M, Steitz JA. Ro small cytoplasmic ribonucleoproteins are a subclass of La ribonucleoproteins: Further characterization of the Ro and La small ribonucleoproteins from uninfected mammalian cells. Mol Cell Biol. 1981;12:1138–49. - PMC - PubMed
    1. Madore SJ, Wieben ED, Pederson T. Eukaryotic small ribonucleoproteins. Anti-La human autoantibodies react with U1 RNA–protein complexes. J Biol Chem. 1984;259:1929–33. - PubMed
    1. Rinke J, Steitz JA. Association of the lupus antigen La with a subset of U6 snRNA molecules. Nucl Acids Res. 1985;13:2617–29. - PMC - PubMed
    1. Pruijn GJM, Slobbe RL, van Venroooij WJ. Structure and function of La and Ro RNPs. Mol Biol Rep. 1990;14:43–48. - PubMed

Publication types