African Swine Fever Virus Gene Research Articles

Promoter analysis of an African swine fever virus gene encoding a putative elongation factor.

Promoter analysis of an African swine fever virus gene encoding a putative elongation factor.

An African Swine Fever Virus Gene with Similarity to the T-Lymphocyte Surface Antigen CD2 Mediates Hemadsorption

An open reading frame, LMW8-DR, in the African swine fever virus (ASFV) genome possesses striking similarity to the lymphocyte membrane antigen CD2. All characterized CD2 domains, including the amino-terminal signal sequence, IgV, hinge, IgC2, stalk, transmembrane, and proline-rich carboxy cytoplasmic domains, are highly conserved in the ASFV gene. Critical residues for the binding of the lymphocyte function-associated antigen (LFA-3) and CD59 and for T-cell activation are also partially conserved. LMW8-DR is actively transcribed in ASFV-infected swine macrophages and Vero cells at late times in the infection cycle and Vero and COS cells transiently expressing the LMW8-DR open reading frame hemadsorbed swine red blood cells. The structural and functional similarities of LMW8-DR to CD2, a protein that is involved in cell-cell adhesion and immune response modulation, suggest a possible role in the pathogenesis of ASFV infection.

Characterization of vaccinia virus gene B12R

We report the characterization of vaccinia virus gene B12R which is predicted to encode a 33K protein with 36% amino acid identity to the serine/threonine protein kinase encoded by vaccinia virus gene B1R. S1 nuclease protection experiments showed that gene B12R is transcribed early during infection from an initiation site 11 bp upstream of the open reading frame (ORF). The gene encodes a 33K polypeptide that is not required for virus replication in tissue culture nor for virus virulence in a murine intranasal model. Expression of the B12R gene in Escherichia coli produced an abundant 33K polypeptide which lacked protein kinase activity under conditions in which the protein kinases encoded by vaccinia virus gene B1R and African swine fever virus gene j9L are active.

An African swine fever virus gene with a similarity to eukaryotic RNA polymerase subunit 6.

An African swine fever virus gene with a similarity to eukaryotic RNA polymerase subunit 6.

Vaccinia virus-mediated expression of african swine fever virus genes

Bacteriophage λ and plasmid clones containing African swine fever virus (ASFV) DNA inserts, which together covered more than 90% of the genome of a Malawi ASFV isolate (LIL 20/1), were transfected into vaccinia virus (VV)-infected cells. Expression of ASFV-encoded proteins was assayed at late times after VV infection by immunoprecipitation of [ 35S]meth ionine-labeled proteins with hyperimmune serum from ASFV-infected pigs, separation of immunoprecipitated proteins by denaturing polyacrylamide gel electrophoresis, and detection by autoradiography. Synthesis of eight additional proteins not observed in control experiments was detected. Seven VV recombinants were constructed, each containing an ASFV DNA insert from a separate bacteriophage λ clone ranging in size from 9 to 15 kb. BSC40 cells were infected with recombinant viruses and expression of ASFV-encoded proteins assayed at early and late times postinfection. Synthesis of additional proteins, not observed in control experiments, was detected by immunoprecipitation with ASFV antiserum both early and late postinfection with two of these recombinants. In these experiments VV promoters were not included upstream of individual ASFV genes.

African Swine Fever Virus Gene Expression in Infected Vero Cells

Polypeptides synthesized in Vero cells infected with African swine fever virus (ASFV) can be divided into early and late classes on the basis of their sensitivity to cytosine arabinoside. As ASFV does not inhibit cell protein synthesis until late in infection, immunoprecipitation was used to identify virus-specific polypeptides. Eighteen early and 15 late polypeptides were detected by polyacrylamide gel electrophoresis. Early polypeptides can be further divided into those which are transiently expressed at early times and the majority which are synthesized throughout infection. In vitro translation products of RNAs from infected cells at different stages of infection were compared with in vivo products after immunoprecipitation. A good correspondence was observed between the in vivo and in vitro patterns. All early RNAs translated in vitro were synthesized in the presence of cytosine arabinoside and cycloheximide and can therefore be classified as immediate early RNAs. Only two polypeptides transcribed from early RNA were not present among late RNA products. No evidence was obtained for a discrete class of delayed early RNAs.