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Biomedical subjects

M Yaniv

Publications and source records attributed to M Yaniv.

At least 73 records · Page 4Linked to original sources
PubMed1997

Hepatocyte nuclear factor 1alpha gene inactivation impairs chromatin remodeling and demethylation of the phenylalanine hydroxylase gene.

Hepatocyte nuclear factor 1 alpha (HNF1alpha) is a homeoprotein that is expressed in the liver, kidney, pancreas, and digestive tract. Its inactivation in mouse resulted in decreased transcription of known target genes such as albumin and alpha1-antitrypsin. In contrast, the phenylalanine hydroxylase (PAH) gene was totally silent and unresponsive to normal inducers like glucocorticoids and cyclic AMP in the liver. DNase I and micrococcal nuclease digestion of liver nuclei showed that HNF1alpha inactivation had drastic effects on the chromatin structure of the PAH regulatory regions. Three DNase I-hypersensitive sites (HSSI, HSSII, and HSSIII), typical of the actively transcribed PAH gene, were undetectable in liver from HNF1alpha-deficient animals. Both HSSII and HSSIII elements harbor HNF1 sites, but only the latter has detectable enhancer activity in transient-transfection assays. In addition, the PAH promoter in livers of HNF1alpha-deficient animals was methylated. These results suggest that HNF1alpha could activate transcription through two mechanisms. One implies participation in the recruitment of the general transcription machinery to the promoter, and the second involves the remodeling of chromatin structure and demethylation that would allow transcription factors to interact with their cognate cis-acting elements.

Animals↗
PubMed1996

Purification and biochemical heterogeneity of the mammalian SWI-SNF complex.

We have purified distinct complexes of nine to 12 proteins [referred to as BRG1-associated factors (BAFs)] from several mammalian cell lines using an antibody to the SWI2-SNF2 homolog BRG1. Microsequencing revealed that the 47 kDa BAF is identical to INI1. Previously INI1 has been shown to interact with and activate human immunodeficiency virus integrase and to be homologous to the yeast SNF5 gene. A group of BAF47-associated proteins were affinity purified with antibodies against INI1/BAF47 and were found to be identical to those co-purified with BRG1, strongly indicating that this group of proteins associates tightly and is likely to be the mammalian equivalent of the yeast SWI-SNF complex. Complexes containing BRG1 can disrupt nucleosomes and facilitate the binding of GAL4-VP16 to a nucleosomal template similar to the yeast SWI-SNF complex. Purification of the complex from several cell lines demonstrates that it is heterogeneous with respect to subunit composition. The two SWI-SNF2 homologs, BRG1 and hbrm, were found in separate complexes. Certain cell lines completely lack BRG1 and hbrm, indicating that they are not essential for cell viability and that the mammalian SWI-SNF complex may be tailored to the needs of a differentiated cell type.

Amino Acid Sequence↗
PubMed1996

The hbrm and BRG-1 proteins, components of the human SNF/SWI complex, are phosphorylated and excluded from the condensed chromosomes during mitosis.

In yeast, the SNF/SWI complex is believed to regulate transcription by locally altering the chromatin structure. At the present time, three human homologues of yeast SNF/SWI proteins have been characterized: hbrm and BRG-1, homologues of SNF2/SWI2, and hSNF5, a homologue of SNF5. We show here that, during mitosis, hbrm and BRG-1 are phosphorylated and excluded from the condensed chromosomes. In this phase of the cell cycle, the level of hbrm protein is also strongly reduced, whereas the level of BRG-1 remains constant. The mitotic phosphorylation of hbrm and BRG-1 is found not to disrupt the association of these proteins with hSNF5 but correlates with a decreased affinity for the nuclear structure in early M phase. We suggest that chromosomal exclusion of the human SNF/SWI complex at the G2-M transition could be part of the mechanism leading to transcriptional arrest during mitosis.

Antibodies↗
PubMed1996

A single element mediates glucocorticoid hormone response of HPV18 with no functional interactions with AP1 or hbrm.

We determined that the human papillomavirus type 18 (HPV18) regulatory region contains one functional GRE sequence that interacts with the glucocorticoid receptor. This sequence conferred a moderate hormonal activation to the HPV18 P105 promoter. Two modulators of glucocorticoid hormone activity, AP1 and hbrm, both involved in P105 transcription, were found not to interfere with this hormonal activation.

Base Sequence↗
PubMed1996

Hepatocyte nuclear factor 1 inactivation results in hepatic dysfunction, phenylketonuria, and renal Fanconi syndrome.

HNF1 is a transcriptional activator of many hepatic genes including albumin, alpha1-antitrypsin, and alpha- and beta-fibrinogen. It is related to the homeobox gene family and is predominantly expressed in liver and kidney. Mice lacking HNF1 fail to thrive and die around weaning after a progressive wasting syndrome with a marked liver enlargement. The transcription rate of genes like albumin and alpha1-antitrypsin is reduced, while the gene coding for phenylalanine hydroxylase is totally silent, giving rise to phenylketonuria. Mutant mice also suffer from severe Fanconi syndrome caused by renal proximal tubular dysfunction. The resulting massive urinary glucose loss leads to energy and water wasting. HNF1-deficient mice may provide a model for human renal Fanconi syndrome.

Animals↗
PubMed1996

The outcome of pregnancy in an immigrant Ethiopian population in Israel.

To study the assumption that obstetric risk factors would change over time in an immigrant Ethiopian population, a retrospective study of the records of singleton births among the Ethiopian immigrants between 1981-88 was undertaken and compared to a similar study of the general population. The Ethiopian immigrants were characterized by pregnancies at age extremes, a greater incidence of preeclampsia and premature deliveries, and a lower incidence of premature rupture of membranes. There was over time an increase in maternal weight gain among the Ethiopians as well as an increase in the incidence of preeclampsia and perinatal mortality.

Adult↗
PubMed1996

Two RAREs and an overlapping CRE are involved in the hepatic transcriptional regulation of the Q10 MHC class I gene.

The Q10 gene is a member of the major histocompatibility complex of the mouse that is expressed in the liver and kidney of the adult. Using transient expression assays, we found that the Q10 promoter was activated by retinoic acid (RA) and exogenous RARs and/or RXRs in a cell type-dependent manner. In addition, the basal activity of the Q10 promoter in HepG2 cells is lowered by expressing a dominant negative form of RARalpha. Incidentally, we have identified two cis-elements which consist of sequences related to retinoic acid response elements (RAREs) and a putative cAMP responsive element (CRE) the sequence of which overlaps one of the RAREs. RAR, RXR, CREB-ATF, and COUP-TF factors bind these elements and/or affect their activity. We also demonstrate that the CRE mediates part of the stimulation induced by activation of the cAMP pathway on the Q10 promoter, the residual activation being mediated by RARs. Our results suggest that Q10 expression in liver depends upon RA and the interaction between nuclear receptors that are expressed in this organ. The overlapping of the CRE with one of the RAREs together with the results of PKA activation also suggest that RA and cAMP signalling pathways are linked.

Journal Article↗
PubMed1995

Control of HPV 18 DNA replication by cellular and viral transcription factors.

Papillomavirus replication in vivo requires the interaction of the virally encoded proteins E1 and E2 with the origin of replication which is localised in the regulatory region (long control region or LCR) of the viral genome. In genital human papillomaviruses (HPVs), the origin overlaps promoter elements of early transcription. In this study, we analysed the replication of HPV18 DNA using the complete LCR containing mutations in transcription regulatory elements. We found that each of the three E2 binding sites proximal to the AT-rich sequence of the origin contributes to the replication rate of DNA, although not identically. In addition, two sequences important for early transcription, an Sp1 binding site and the TATA box, were also found to play a role in replication. In contrast, two AP1 binding sites required for the enhancer-mediated activation of early transcription did not affect the replication, while other upstream sequences in the LCR did contribute to the replication efficiency. Our results indicate that besides a core origin of replication containing an AT-rich sequence and three E2 binding sites, auxiliary elements affect HPV18 DNA replication in the context of the full length LCR, some of which are important for transcription.

Adenovirus E2 Proteins↗
PubMed1995

A human protein with homology to Saccharomyces cerevisiae SNF5 interacts with the potential helicase hbrm.

In yeast, the SNF/SWI complex is involved in transcriptional activation of several inducible promoters, possibly by causing a local modification of the chromatin structure. Recently, two human homologues of the SNF2/SWI2 protein have been isolated, hbrm and BRG-1. In addition, a complex containing one of the SNF2/SWI2 homologues and having an in vitro activity similar to the yeast complex has been partially purified from HeLa cells. Here we describe the characterization of a cDNA encoding a human nuclear protein containing a large domain of homology with SNF5, another member of the yeast SNF/SWI complex. This protein can be co-immunoprecipitated with hbrm and the interaction between the two proteins is dependent on the region conserved between the human and the yeast SNF5. These findings suggest that the cDNA we have cloned encodes one of the members of the human SNF/SWI complex.

Amino Acid Sequence↗
PubMed1995

The bovine papillomavirus 1 E2 protein contains two activation domains: one that interacts with TBP and another that functions after TBP binding.

The E2 transactivator of bovine papillomavirus type-1 is unable to activate minimal promoters in vivo that contain only E2 binding sites and a TATA box. This block can be overcome by over-expression of human TATA binding protein (TBP) or by the addition of either SP1 binding sites or an initiator element to the promoter, suggesting that the binding of TFIID may normally be a rate-limiting step for activation by E2. Surprisingly, purified E2 and TBP bind co-operatively to DNA in vitro when the sites are closely spaced. E2 does not affect the on rate of association but reduces the off rate. The E2 region responsible for this effect is located in the hinge region that links the classic transactivation and DNA binding domains. We demonstrate that the TBP stabilizing domain contributes in vivo to co-operativity with co-expressed TBP and to activation of the major late minimal promoter (MLP) containing E2 sites. In contrast, promoters with SP1 sites are activated to wild-type levels by such a mutant. This promoter specificity is also evident in vitro. A truncated E2 mutant, lacking the classic transactivation domain but containing the TBP stabilizing domain, stimulates transcription of the MLP in vitro, but does not activate promoters with SP1 sites. In conclusion, our results show that the E2 transactivation domain has a modular structure. We have identified one domain which probably acts at an early step in the assembly of the pre-initiation complex and which is involved in reducing the dissociation rate of bound TBP in vitro. The classic N-terminal activation domain of E2 might affect one or several step(s) in the assembly of the preinitiation complex occurring after the binding of TFIID.

Adenoviridae↗
PubMed1995

A c-Jun dominant negative mutant protects sympathetic neurons against programmed cell death.

Sympathetic neurons depend on nerve growth factor (NGF) for survival and die by apoptosis in its absence. We have investigated the pattern of expression of the Jun and Fos family of transcription factors in dying sympathetic neurons using antibodies specific for each family member. When sympathetic neurons are deprived of NGF, the level of c-Jun protein significantly increases, whereas the levels of the other members of the Jun and Fos family remain relatively constant. c-Jun also becomes more phosphorylated, probably on its amino terminal transactivation domain. When microinjected into sympathetic neurons, an expression vector for a c-Jun dominant negative mutant protects them against NGF withdrawal-induced death, indicating that AP-1 activity is essential for neuronal cell death. Furthermore, overexpression of the full-length c-Jun protein is, in itself, sufficient to induce apoptosis in sympathetic neurons.

Animals↗
PubMed1994

Mouse JunD negatively regulates fibroblast growth and antagonizes transformation by ras.

As NIH 3T3 fibroblasts become quiescent, the level of c-Jun protein decreases while JunD accumulates. When resting cells are stimulated with fresh serum, nuclear-localized JunD is rapidly degraded, followed by resynthesis of both c-Jun and JunD later in G1. Overexpression of JunD results in slower growth and an increase in the percentage of cells in G0/G1 while c-Jun overexpression produces larger S/G2 and M phase populations. In addition, JunD partially suppresses transformation by an activated ras gene whereas c-Jun cooperates with ras to transform cells. These data indicate that two closely related transcription factors can function in an opposing manner.

Amino Acid Sequence↗
PubMed1994

Cooperativity in vivo between the E2 transactivator and the TATA box binding protein depends on core promoter structure.

The E2 transactivator protein of bovine papillomavirus 1 (BPV-1) can strongly stimulate complex promoters such as that of the herpes simplex virus thymidine kinase gene but does not efficiently activate minimal promoters that only contain E2 binding sites and a TATA box. Here we show that overexpression of the human, but not yeast, TATA box binding protein (TBP) in transfection experiments overcomes this block and enables E2 to activate a minimal TATA box-containing promoter. This suggests that recruitment of the TFIID complex to such promoters is normally a rate limiting step for transcriptional activation by E2 in vivo. In contrast, minimal promoters that contain an initiator element in addition to a TATA box are efficiently activated by E2 on its own and this activation is only moderately enhanced by TBP overexpression. In such E2-responsive promoters the TATA box or initiator can be functionally replaced by SP1 binding sites. Both the initiator binding protein, TFII-I, and SP1 have been found to interact physically with components of the TFIID complex. Since either TBP overexpression or the presence of an initiator or SP1 binding sites can increase activation by E2, it seems likely that the principal role of the E2 activation domain is to affect a step in the formation of the transcription initiation complex that occurs after TFIID has bound to the promoter. Sequential action of transcription factors, such as TFII-I, SP1 and E2, may be one type of mechanism underlying the widely observed phenomenon of transcriptional synergy.

Adenoviridae↗
PubMed1994

The E2 transcriptional repressor can compensate for Sp1 activation of the human papillomavirus type 18 early promoter.

The E6/E7 early promoter (P105) of genital human papillomavirus type 18 contains binding sites for the viral regulator E2, tandemly repeated and closely flanked by two crucial promoter elements; the TATA box downstream and an Sp1 binding site upstream. We showed that binding of purified E2 and Sp1 proteins in vitro to their neighboring sites is mutually exclusive and that Sp1 is displaced by E2. However, this displacement did not result in repression of P105 transcription. In contrast, binding of E2 to its site overlapping the Sp1 binding site activated transcription of P105 derivatives lacking the E2 site most proximal to the TATA box. Surprisingly, a truncated form of E2, deleted of part of the transactivation domain and known as the E2 transcriptional repressor, as well as the E2 DNA-binding domain alone also supported transcription of these P105 derivatives. In the context of P105, the viral E2 protein can thus activate P105 transcription in place of Sp1, even in the absence of its transactivation domain.

Base Sequence↗
PubMed1994

The bovine papillomavirus type 1 (BPV1) replication protein E1 modulates transcriptional activation by interacting with BPV1 E2.

The study of bovine papillomavirus type 1 (BPV1) DNA replication has shown that E1 and E2 are the only viral proteins required for this process. Both E1 and E2 interact with the viral origin of replication (ori). The BPV1 E2 protein is also a well-characterized transcriptional regulator. We show in this report that E1 can modulate transcription by interactions with E2. At low concentrations, E1 enhanced the E2-mediated transactivation of heterologous promoters containing the BPV1 ori by promoting cooperative binding of both E1 and E2 to the DNA. In contrast, in the presence of excess E1, transactivation by E2 is repressed. This last process, however, does not require cooperative DNA binding of the two proteins. These results imply that the balance between these two distinct types of interaction is crucial both for control of replication and for early viral transcription.

Base Sequence↗
PubMed1993

Functional interference between the Spi-1/PU.1 oncoprotein and steroid hormone or vitamin receptors.

The Spi-1/PU.1 protein is an Ets-related transcription factor, whose overexpression is a consequence of SFFV integration in Friend erythroleukemic cells. We present evidence that Spi-1/PU.1 can specifically repress the glucocorticoid-induced activation of promoters carrying a glucocorticoid response element (GRE). Conversely, the glucocorticoid receptor (GR) represses Spi-1/PU.1-mediated transcriptional activation in the presence of hormone. Spi-1/PU.1 also antagonized activation by other nuclear receptors, such as the thyroid hormone or the retinoic acid receptors, in several cell lines, including K562 erythroleukemic cells. These observations suggest that accumulation of the Spi-1/PU.1 protein may interfere with the action of hormones in the erythrocyte differentiation pathway.

Base Sequence↗