The nucleotide sequence of the transforming BglII-H fragment of adenovirus type 7 DNA

Autoregulation of the adenovirus EIA gene involves its constitutive expression and positively and negatively regulated transcription. Dissection of this process will identify basal-level cis elements and autoregulatory targets of the ElA promoter and functional domains within the transacting ElA gene products. In this report, the DNA sequence of the human subgroup B adenovirus type 3 (Ad3) ElA gene is presented and compared with that of the EIA genes of similar and distantly related human adenoviruses. The cDNA forms of the Ad3 ElA gene, corresponding to two major early mRNA species, are cloned, sequenced, and subcloned into plasmid expression vectors. Cotransfections of cell cultures are performed with Ad5 or Ad3 ElA gene expression plasmids and a reporter gene under control of the Ad5 or Ad3 ElA promoter. The Ad5 and Ad3 ElA promoters are similarly repressed by either serotype's 12S cDNA gene products. The Ad3 ElA promoter responds much more strongly than the AdS EIA promoter to transactivation by 13S cDNA gene products. In contrast, the 13S cDNA gene of Ad5 has greater transactivation activity than that of Ad3. Experiments with missense mutations

Section: Methodssupporting

confidence: 80%

Section: Resultsmentioning

confidence: 57%

Functional diversity of E1A gene autoregulation among human adenoviruses

Cogan

Jones

Hall

et al. 1992

“…1) requires isolation of an appropriate Ad7a DNA fragment (viral backbone) which is deleted of left-end elements required for virus replication and packaging. Generating such a fragment from Ad7a is complicated by the fact that the majority of the Ad7a DNA sequence has not been determined (7,8,15). By analyzing the available Gomen strain Ad7 left-end sequence, we identified a battery of restriction endonucleases which were able to digest Ad7 DNA at an appropriate site at the left end.…”

Section: First-generation Adenovirus Vectors Include Virus Constructsmentioning

confidence: 99%

Construction of an adenovirus type 7a E1A- vector

Abrahamsen

Kong

Mastrangeli

et al. 1997

A strategy for constructing replication-defective adenovirus vectors from non-subgroup C viruses has been successfully demonstrated with adenovirus type 7 strain a (Ad7a) as the prototype. An E1A-deleted Ad7a reporter virus expressing the chloramphenicol acetyltransferase (CAT) gene from the cytomegalovirus promoter enhancer was constructed with DNA fragments isolated from Ad7a, an Ad7a recombination reporter plasmid, and the 293 cell line. The Ad7a-CAT virus particle transduces A549 cells as efficiently as Ad5-based vectors. Intravenous infections in a murine model indicate that the Ad7a-CAT virus infects a variety of tissues, with maximal levels of CAT gene expression found in the liver. The duration of Ad7a-CAT transgene expression in the liver was maximally maintained 2 weeks postinfection, with a decline to baseline activity by the week 4 postinfection. Ad7a-CAT represents the first example of a non-subgroup C E1A ؊ adenovirus gene transfer vector.

“…It is deduced from these observations that EBB gene products play an important role in the transformation of cells. DNA sequence data of the EBB gene reveal that there are at least two polypeptides encoded by EBB (19,000-molecular-weight [19K] and 54K polypeptides in Adl2) (3,4,16,25,29). The smallest Adl2 DNA fragment capable of transforming rodent cells completely is the HindIII-G (0 to 6.8 map units) (22).…”

mentioning

confidence: 99%

“…Thus, transcriptional control is not affected by the mutation. 4. DNA and the corresponding amino acid sequences of the mutated viruses.…”

mentioning

confidence: 99%

Isolation of transformation-defective, replication-nondefective early region 1B mutants of adenovirus 12

Fukui¹,

Saito²,

Shiroki³

et al. 1984

We isolated three adenovirus 12 early region 1B mutants (in2O5B, in2O5C, and d1205) by ligation of the cleaved DNA-protein complex and transfection of human embryo kidney cells with the ligation products. These mutants could replicate efficiently in human embryo kidney or KB cells but showed markedly reduced transforming capacities both in vitro and in vivo. In cells infected with the mutants, the early region 1B gene was transcribed efficiently. In cells infected with in2O5B, the products corresponding to the early region lB-coded 19,000-molecular-weight polypeptide was detected by in vitro translation but not immunoprecipitated extract of labeled cells. In cells infected with in2O5C or d1205, the products corresponding to the same polypeptide were not detected by either in vitro translation or immunoprecipitation of labeled cell extracts. The results suggest that the 19,000-molecular-weight polypeptide encoded by early region 1B is required for cell transformation but not for viral propagation.