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Publications de l’équipe
Plasticité épigénétique et polarité de l’embryon
Année de publication : 1989
P F Lambert, N Dostatni, A A McBride, M Yaniv, P M Howley, B Arcangioli (1989 Jan 1)
Functional analysis of the papilloma virus E2 trans-activator in Saccharomyces
cerevisiae.
Genes & development : 38-48
Résumé
The papilloma virus E2 transcriptional trans-activator is representative of a class of
transcriptional modulators that activate transcription through direct binding to cis-acting
DNA sequences. In this study we measured the capacity for this mammalian virus factor to
function in Saccharomyces cerevisiae. When expressed in the yeast, the bovine papilloma
virus E2 trans-activator could stimulate transcription from a yeast promoter having E2 DNAbinding sites present in cis. Whereas a single E2 DNA-binding site was sufficient for transactivation, a strong cooperative effect was observed with two E2 DNA-binding sites. The level
of trans-activation was dependent on the position of the E2 DNA-binding sites in relation to
the yeast promoter, with the maximal effect demonstrated when the binding sites were
positioned upstream. Deleted E2 proteins, lacking part of the trans-activation or DNA-binding
domains, failed to activate transcription in yeast, similar to their behavior in mammalian
cells. Replacement of the amino-terminal region of the E2 trans-activation domain with a
synthetic amphipathic helix partially restored the trans-activation function; however, it did
not result in a molecule that exhibited cooperativity between neighboring E2 DNA-binding
sites.
Année de publication : 1988
N Dostatni, M Yaniv, O Danos, R C Mulligan (1988 Dec 1)
Use of retroviral vectors for mapping of splice sites in cottontail rabbit
papillomavirus.
The Journal of general virology : 3093-100
Résumé
Cottontail rabbit papillomavirus (CRPV) genomic sequences coding for virus early functions
were introduced into a retroviral vector in order to produce cDNAs of the viral early region.
Two constructs differing in the length of control sequences preceding the E6 open reading
frame were transfected into Psi-2 cells and the released retroviral stock was used to infect
NIH3T3 cells. The proviral sequences were rescued from antibiotic G418-resistant virusinfected cells after fusion with Cos cells, amplified as plasmids in Escherichia coli and
analysed. Nucleotide sequencing showed that the splicing signals used in the construct
containing only the early coding region are the same as in CRPV-expressing tumours, linking
the beginning of E1 to the middle of E2. On the other hand, in a construct including most of
the long control region a splice donor site located in the 5′ end of this region, at position
7810, was very efficiently used, totally excluding the use of the donor site at position 1371.
None of the constructs containing CRPV sequences transcribed from Moloney murine
INSTITUT CURIE, 20 rue d’Ulm, 75248 Paris Cedex 05, France | 1
Publications de l’équipe
Plasticité épigénétique et polarité de l’embryon
leukaemia virus promoter was able to transform mouse fibroblasts after DNA transfection.
N Dostatni, F Thierry, M Yaniv (1988 Dec 1)
A dimer of BPV-1 E2 containing a protease resistant core interacts with its DNA
target.
The EMBO journal : 3807-16
Résumé
The E2 proteins encoded by papillomaviruses interact with the viral DNA to regulate its
transcription. In the present study, we have constructed bacterial vectors expressing the fulllength or N-terminal truncated BPV-1 E2 proteins under the control of an inducible promoter.
By UV cross-linking experiments we show that a dimer of the intact or truncated E2 protein
interacts with a single palindromic site ACCGNNNNCGGT. The DNA-binding domain of E2 can
be reduced to a small protease resistant core. Methylation interference studies show that
this C-terminal domain interacts with the major groove of the DNA by contacting two
consecutive guanine residues in both halves of the palindrome. Although one binding site is
sufficient for high affinity binding in vitro or in vivo, two E2 binding sites are required for
transcriptional activation in eukaryotic cells.
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