A unique amino-terminal sequence predicted for the chicken proto-ets protein

Avian cellular sequences homologous to the ets domain of the avian leukemia retrovirus, E26, have been characterized, and shown to contain nine discrete regions within a single locus of about 60 kb. The structure of the viral homologous and nonhomologous domains of this chicken ets-1 gene is characterized and seen to define the unique amino acid sequences of the ets protein. We have isolated and sequenced an avian ets-1 cDNA clone obtained from chicken thymus. This cDNA clone contains an open reading frame (ORF) encoding the normal cellular product of 441 amino acids. This product is significantly smaller than that encoded by the v-ets domain of the E26 transforming protein, p135, which contains 491 amino acids. The open reading frame predicted by our sequence data results in a protein calculated to be 50 kDa, which is slightly smaller than that actually observed in chicken cells. The presence of termination codons 5' to this ORF demonstrates that the cDNA characterized contains the entire coding region for the chicken ets gene.     

The specificity of the disease induced by defective murine retroviruses containing abl, fos, or ha-ras is usually not determined by their LTR

The long terminal repeats (LTR) of the defective murine sarcoma viruses (MSV) containing v-abl, v-Ha-ras, or v-fos were exchanged for LTRs from other retroviruses having different tissue tropism. The new chimeric MSV were found to induce the same diseases as the parental viruses, indicating that sequences outside the LTR, most likely those of the oncogene, are responsible for the disease specificity of these defective MSV.     

Ig/EBP-1: a ubiquitously expressed immunoglobulin enhancer binding protein that is similar to C/EBP and heterodimerizes with C/EBP

We report the isolation and characterization of cDNA clones that encode a protein with the same DNA binding specificity as the immunoglobulin heavy chain enhancer binding protein E (muEBP-E). We call the gene encoding this protein Ig/EBP-1. A fusion protein encoded by the cDNA binds specifically to muEBP-E-binding sites (E sites) in both the IgH enhancer and the VH1 promoter. Sequence analysis reveals that Ig/EBP-1 is a member of the "basic-zipper" family of DNA-binding proteins that are characterized by basic regions and heptad repeats of leucine residues. Among known family members, Ig/EBP-1 demonstrates highest homology to C/EBP throughout the DNA-binding domain and leucine repeat region. Ig/EBP-1 and C/EBP have highly overlapping binding specificities; both cloned proteins bind to the IgH enhancer and the VH1 promoter E sites, and Ig/EBP-1 binds to previously characterized C/EBP binding sites in the Rous sarcoma virus (RSV) LTR and the murine albumin promoter. Consistent with their homology in the leucine repeat region, Ig/EBP-1 and C/EBP form heterodimers; Ig/EBP-1 is the first member of this family that has been found to heterodimerize with the well-characterized C/EBP. Ig/EBP-1 mRNA is present in all tissues and cell lines examined, although its levels vary almost 20-fold from different sources, with highest levels in early B cells. In tissues where Ig/EBP-1 and C/EBP are both present, heterodimers may be functionally important. The presence of Ig/EBP-1 in fibroblasts and other tissues where C/EBP is not expressed suggests that Ig/EBP-1 may be functionally important for the activity of the RSV enhancer in these cell types. Finally, elevated expression of Ig/EBP-1 in early B cells may explain in part the enhancer-independent activity of VH promoters early in B-cell development.     

Identification of Residues within the Herpes Simplex Virus Type 1 Origin-Binding Protein That Contribute to Sequence-Specific DNA Binding

Gene UL9 of herpes simplex virus type 1 encodes an 851-amino-acid protein which is essential for viral DNA synthesis and functions as a sequence-specific origin-binding protein and DNA helicase. We generated monoclonal antibodies against purified UL9 protein and identified one such antibody (MAb 13924) that can block the interaction of the UL9 C-terminal DNA-binding domain (amino acids 534–851) with its recognition sequence. MAb 13924 interacted with immobilized peptides containing residues 780–786 of UL9. Although the corresponding region of the homologous protein encoded by varicella-zoster virus differs at only a single position it was not recognized by MAb 13924. Site-directed mutagenesis experiments confirmed that residues within this region contribute to the epitope recognized by MAb 13924 and may be involved in sequence-specific DNA binding. In addition, all eight lysine residues within the DNA-binding domain were separately changed to alanine and the DNA-binding properties of the mutated proteins were examined. The results showed that lysine residues that are located close to the peptide recognized by MAb 13924 or lie within the region of the DNA-binding domain most highly conserved among homologous alphaherpesvirus proteins play a role in sequence-specific DNA binding. Moreover, alteration of a lysine residue 18 amino acids from the recognized peptide prevented the interaction of MAb 13924 with the UL9 C-terminal DNA-binding domain. Three helical segments are predicted to occur within the region containing mutations that affect sequence-specific binding and interaction with MAb 13924.     

ski can cause selective growth of skeletal muscle in transgenic mice

We have created several lines of mice that contain a truncated chicken c-ski cDNA linked to an MSV LTR promoter. Adult mice from three independent lines show large increases in skeletal muscle. All three lines of mice express high levels of c-ski mRNA and protein in skeletal muscle. All other tissues examined show little or no expression of the c-ski transgene. The muscles of one of the three lines were examined in more detail. Type II fast fibers undergo selective hypertrophy in affected muscles of this line.