Regulation of adenovirus transcription by an Ela gene in microinjected Xenopus laevis oocytes

Journal Article · Thu Dec 01 04:00:00 EST 1983 · Mol. Cell. Biol.; (United States)

DOI:https://doi.org/10.1128/MCB.3.12.2131· OSTI ID:5942093

Jones, N C; Richter, J D; Weeks, D L; Smith, L D

The regulation of adenovirus type 5 gene expression by the E1a gene product was examined in microinjected Xenopus laevis oocytes. Chimeric genes were constructed which included the promoter region of early adenovirus type 5 gene 3 and the structural sequence which codes for the bacterial enzyme chloramphenicol-3-O-acetyltransferase (CAT). A plasmid containing this chimeric gene as well as plasmids containing the E1a gene were coinjected into oocyte nuclei. The presence of the E1a gene was shown to increase CAT activity by up to 8.5-fold over basal levels. Synthesis of the functional product from the E1a gene requires the removal of intron sequences by RNA splicing. The E1a gene and a derivative that precisely lacks the intron were equally effective in increasing CAT activity, suggesting that splicing of the primary E1a transcript is efficiently accomplished in the oocyte nucleus. This was confirmed by directly examining the E1a mRNAs by the S1 mapping procedure. A protein extract from adenovirus type 5-infected HeLa cells enriched for the E1a protein may supplant the E1a plasmid in enhancing CAT activity. Synthesis of the CAT enzyme after gene injection is invariant in oocytes from the same frog, but oocytes from different frogs show a high degree of variability in their ability to synthesize the CAT enzyme. Microinjected X. laevis oocytes appear to be an extremely useful system to study the effects of protein elements on transcription.

• Research Organization: Dept. of Biological Sciences, Purdue Univ., West Lafayette, IN 47907
• OSTI ID: 5942093
• Journal Information: Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 3:12; ISSN MCEBD
• Country of Publication: United States
• Language: English

Related Subjects

550200 -- Biochemistry
550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
ACETYL RADICALS
ACYL RADICALS
ADENOVIRUS
AMPHIBIANS
ANIMALS
ANTI-INFECTIVE AGENTS
ANTIBIOTICS
AQUATIC ORGANISMS
BACTERIA
BIOTECHNOLOGY
CELL CONSTITUENTS
CHIMERAS
CHLORAMPHENICOL
CONTROL
DNA
DNA SEQUENCING
DRUGS
ENZYME INDUCTION
ENZYMES
FROGS
GAMETES
GENE REGULATION
GENETIC CONTROL
GENETIC ENGINEERING
GERM CELLS
HELA CELLS
HYBRIDIZATION
MICROORGANISMS
MOSAICISM
NUCLEIC ACIDS
ONCOGENIC VIRUSES
ORGANIC COMPOUNDS
OVA
PARASITES
PEST CONTROL
PLASMIDS
RADICALS
RECOMBINANT DNA
SENSITIVITY
STRUCTURAL CHEMICAL ANALYSIS
TRANSCRIPTION
TRANSFERASES
VERTEBRATES
VIRUSES