Structure and organization of the gene coding for the DNA binding protein of adenovirus type 5

Abstract

We have determined the nucleotide sequence of a region of adenovirus type 5 (Ad5) DNA located between map positions 61.7 and 71.4, which covers the gene form the 72 kD DNA binding protein (DBP) and the sequence encoding the amino-terminal part of the 100 kD protein. Sequence analysis of cDNA copies of DBP mRNA revealed the existence of two abundant species of spliced mRNA molecules. One species consists of two short leader sequences from positions 75.2 (67 and 68 nucleotides long) and 68.8 (77 nucleotides long), respectively, and the main body of the RNA molecules. The other species contains only the leader sequence from position 75.2 and the main body. The amino acid sequence of DBP is encoded entirely by a long open reading frame of 1587 nucleotides in the main body of DBP mRNA. From the nucleotide sequence of the DBP gene it can be derived that DBP contains 529 amino acid residues and has an actual molecular weight of 59,049 daltons. The sites of mutation in the mutants H5hr404 and H5ts125 were determined at the nucleotide level. Single nucleotide alterations were detected in H5hr404 and H5ts125 in the sequences corresponding to the amino-terminal part and the carboxy-terminal part of DBP, respectively. The implications of these mutations are discussed.

mRNA polyadenylate-binding protein: gene isolation and sequencing and identification of a ribonucleoprotein consensus sequence.

“…as regions of similar general sequence character are found in other single-stranded-nucleic acid-binding proteins (15,29). It is possible that the high affinity and binding specificity of the protein for the poly(A) tail are a consequence of an interaction involving these two distinct domains.…”

“…The carboxy third of the molecule, separated from the rest of the protein by a segment of eight consecutive alanine residues, contains a segment (amino acids 426 to 489) of very high proline content and an overall positive charge due to the absence of negatively charged amino acids. Regions rich in proline and positively charged amino acids have been identified in other single-stranded-nucleic acid-binding proteins, such as the nucleocapsid protein of Semliki Forest virus (15) and the adenovirus type 5 single-stranded-DNA-binding protein (29). The amino-terminal half of the molecule has a very different structure.…”

p53 promotes adenoviral replication and increases late viral gene expression

“…We then went on to test whether cell death is linked to virus replication using the temperature-sensitive (ts) virus ts125, which is unable to replicate its DNA at 39.51C (Williams et al, 1974) due to a defect in the DBP (Kruijer et al, 1981), but is replication competent at the permissive temperature of 32.51C. A549 cells were infected with wtAd5 or ts125 and incubated at either 39.5 or 32.51C for 4 days and cell viability determined.…”

Cloned Human snRNP Proteins B and B′ Differ Only in Their Carboxy-Terminal Part

“…The nucleic acid binding property of the proteins B and B' can in part be accounted for by the relatively high content of positively charged amino acids (12%), in comparison to negatively charged residues (5%). Also, proline-rich domains have been described in nucleic acid binding proteins (Garoff et al, 1980;Kruijer et al, 1981).…”