109 research outputs found
Human Papillomaviruses in Buschke-Lowenstein Tumors: Physical State of the DNA and Identification of a Tandem Duplication in the Noncoding Region of a Human Papillomavirus 6 Subtype
Six Buschke-Löwenstein tumors, i.e., highly differentiated squamous cell tumors of the genital region, were shown to contain human papillomavirus 6 (HPV 6) or HPV 11 genomes. The viral DNA was found in an episomal state, including a very small fraction of circular oligomers. HPV 6a and HPV 6d genomes were cloned from two of the tumors. Comparison with HPV 6b, cloned from a benign genital wart (E. -M. de Villiers, L. Gissmann, and H. zur Hausen, J. Virol. 40:932-935, 1981) by restriction mapping and partial sequence analysis, revealed a very high degree of homology with the different HPV 6 subtypes. A tandem duplication of 459 base pairs within the noncoding region of the genome was found in the new subtype HPV 6d. This structural rearrangement in a region containing the putative control elements for early gene transcription might influence the biological potential of that virus. No evidence for rearrangement of this region was found in the HPV DNA from the five other tumors
Reporter constructs with low background activity utilizing the cat gene
Reporter plasmids utilizing the cat gene for the analysis of promoter and enhancer sequences in vertebrate cells, were constructed. These plasmids minimize the background of transcription derived from cryptic promoters or cryptic regulatory elements within the vecto
Extinction of tyrosine aminotransferase gene activity in somatic cell hybrids involves modification and loss of several essential transcription factors
Extinction is defined as the loss of cell type-specific gene expression that occurs in somatic cell hybrids
derived by fusion of cells with dissimilar phenotypes. To explore the basis of this dominant-negative
regulation, we have studied the activities of the control elements of the liver-specific gene encoding tyrosine
aminotransferase (TAT) in hepatoma/fibroblast hybrid crosses. We show that extinction in complete somatic
cell hybrids is accompanied by the loss of activity of all known cell type-specific control elements of the TAT
gene. This inactivity is the result of first, lack of expression of genes coding for the transcriptional activators
HNF4 and HNF3[~ and HNF33,, which bind to essential elements of the enhancers; and second, loss of in vivo
binding and activity of ubiquitous factors to these enhancers, including CREB, which is the target for
repression by the tissue-specific extinguisher locus TSE1. Complete extinction of TAT gene activity is
therefore a multifactorial process affecting all three enhancers controlling liver-specific and hormone-inducible
expression. It results from lack of activation, rather than active repression, and involves both
post-translational modification and loss of essential transcriptional activators
Extinction of gene expression in somatic cell hybrids. a reflection of important regulatory mechanisms?
Extinction in somatic cell hybrids is a multifactorial process that leads to loss of cell-type-specific gene expression. The underlying mechanisms are thought to mirror, at least in part, the repertoire of regulatory mechanisms controlling mammalian cell differentiation
Role of cyclic AMP in the control of cell-specific gene expression
Genes have to be expressed in specific cell types at appropriate times of development dependent on external signals. cAMP signaling occurs in all cells, thus raising the question of how this signal transduction pattern is integrated into mechanisms determining cell-specific gene expression. We have analyzed expression of the tyrosine aminotransferase gene as a model to study the basis of this cell type specificity of hormone induction. We found that cell-type-specific expression is generated by combined action of cAMP signal-dependent and liver cell-specific transcription factors. The interdependence of the cAMP response element and an element determining liver cell specificity enables a gene to respond to an ubiquitous signal in a cell-specific manner
Constancy of Expression of the Protein Kinase A Regulatory Subunit Rla in Hepatoma Cell Lines of Different Phenotypes
Multiple mRNA isoforms of the transcription activator protein CREB
We have characterized cDNA clones representing mouse
CREB (cyclic AMP responsive element binding protein)
mRNA isoforms. These include CREBA and CREBa, of
which the rat and human homologues have been
previously identified. Both encode proteins with CREbinding
activity and identical transactivation potential.
The additional CREB mRNA isoforms potentially encode
CREB related proteins. From the structural organization
of the mouse CREB gene we conclude that the multiple
transcripts are generated by alternative splicing. Furthermore
we show that specific CREB mRNA isoforms are
expressed at a high level in the adult testis. Expression
of these isoforms is induced after commencement of
spermatogenesis. In situ hybridization suggests that this
expression occurs predominantly in the primary spermatocytes.
Comparison of the CREB gene with the recently
isolated CREM (cAMP responsive element modulator)
cDNAs illustrates that the two genes have arisen by gene
duplication and have diverged to encode transcriptional
activators and repressors of the cAMP signal transduction
pathway
A Very Strong Enhancer Is Located Upstream of an Immediate Early Gene of Human Cytomegalovirus
A strong transcription enhancer was identified in the genomic DNA (235 kb) of human cytomegalovirus (HCMV), a ubiquitous and severe pathogen of the herpesvirus group. Cotransfection of enhancerless SV40 DNA with randomly fragmented HCMV DNA yielded two SV40-HCMV recombinant viruses that had incorporated overlapping segments of HCMV DNA to substitute for the missing SV40 enhancer. Within HCMV, these enhancer sequences are located upstream of the transcription initiation site of the major immediate-early gene, between nucleotides -118 and −524. Deletion studies with the HCMV enhancer, which harbors a variety of repeated sequence motifs, show that different subsets of this enhancer can substitute for the SV40 enhancer. The HCMV enhancer, which seems to have little cell type or species preference, is severalfold more active than the SV40 enhancer. It is the strongest enhancer we have analyzed so far, a property that makes it a useful component of eukaryotic expression vectors
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