scholarly journals Sequence Elements Necessary for Transcriptional Activation of BAD1 in the Yeast Phase of Blastomyces dermatitidis

2004 ◽  
Vol 3 (3) ◽  
pp. 785-794 ◽  
Author(s):  
Peggy J. Rooney ◽  
Bruce S. Klein
Keyword(s):  
Transcriptional Activation ◽  
Histoplasma Capsulatum ◽  
Upstream Region ◽  
Blastomyces Dermatitidis ◽  
Specific Gene ◽  
Specific Expression ◽  
Sequence Elements ◽  
Putative Transcription Factor ◽  
Yeast Phase ◽  
Responsive Element

ABSTRACT Blastomyces dermatitidis is a dimorphic fungal pathogen that converts from mycelia or conidia to a host-adapted yeast morphotype upon infection. Conversion to the yeast form is accompanied by the production of the virulence factor BAD1. Yeast-phase-specific expression of BAD1 is transcriptionally regulated, and its promoter shares homology with that of the yeast-phase-specific gene YPS3 of Histoplasma capsulatum. Serial truncations of the BAD1 upstream region were fused to the lacZ reporter to define functional areas in the promoter. Examination of P BAD1 -lacZ fusions in B. dermatitidis indicated that BAD1 transcription is upregulated in the yeast phase. The 63-nucleotide box A region conserved in the YPS3 upstream region was shown to be an essential component of the minimal BAD1 promoter. A matched P YPS3 -lacZ construct indicated that this same region was needed for minimal YPS3 promoter activity in B. dermatitidis transformants. Reporter activity in H. capsulatum transformants similarly showed a requirement for box A in the minimal BAD1 promoter. Several putative transcription factor binding sites were identified within box A of BAD1. Replacement of two of these predicted sites within box A—a cAMP responsive element and a Myb binding site—sharply reduced transcriptional activity, indicating that these regions are critical in dictating the yeast-phase-specific expression of this crucial virulence determinant of B. dermatitidis.

scholarly journals Sequence-specific activation of transcription by adenovirus EIa products is observed in HeLa cells but not in 293 cells.

1986 ◽  
Vol 6 (1) ◽  
pp. 201-208 ◽  
Author(s):  
T Leff ◽  
P Chambon
Keyword(s):  
Hela Cells ◽  
Upstream Region ◽  
Gene Products ◽  
Promoter Regions ◽  
Chimeric Promoter ◽  
293 Cells ◽  
Activation Of Transcription ◽  
Upstream Promoter ◽  
Sequence Elements ◽  
Responsive Element

The adenovirus EIa gene products activate transcription from the viral EIII and EIIaE promoters. We studied the mechanism of this stimulation by constructing a series of chimeric promoter recombinants containing the upstream regions of the EIII and EIIaE promoters linked to the TATA box-start-site regions of the viral major late and EIIa late promoters. By introducing these recombinants into HeLa cells together with recombinants producing the EIa gene products, we demonstrated that the induction of EIII and EIIaE transcription by EIa 13S and 12S mRNA products is dependent on sequences located in the upstream region (approximately -40 to -250) of these promoters. In addition, we showed that the major late and EIIa late upstream promoter regions do not contain such EIa-responsive sequence elements. In contrast, after transfection of these chimeric promoter recombinants into 293 cells (which constitutively express the EIa proteins), we found that their relative levels of transcription are similar and markedly different from those observed when they are cotransfected into HeLa cells with EIa protein-producing recombinants. We conclude that the efficiency of transcription from a given promoter in 293 cells is not necessarily related to the presence of a specific EIa-responsive element.

Sequence-specific activation of transcription by adenovirus EIa products is observed in HeLa cells but not in 293 cells

1986 ◽  
Vol 6 (1) ◽  
pp. 201-208
Author(s):  
T Leff ◽  
P Chambon
Keyword(s):  
Hela Cells ◽  
Upstream Region ◽  
Gene Products ◽  
Promoter Regions ◽  
Chimeric Promoter ◽  
293 Cells ◽  
Activation Of Transcription ◽  
Upstream Promoter ◽  
Sequence Elements ◽  
Responsive Element

The adenovirus EIa gene products activate transcription from the viral EIII and EIIaE promoters. We studied the mechanism of this stimulation by constructing a series of chimeric promoter recombinants containing the upstream regions of the EIII and EIIaE promoters linked to the TATA box-start-site regions of the viral major late and EIIa late promoters. By introducing these recombinants into HeLa cells together with recombinants producing the EIa gene products, we demonstrated that the induction of EIII and EIIaE transcription by EIa 13S and 12S mRNA products is dependent on sequences located in the upstream region (approximately -40 to -250) of these promoters. In addition, we showed that the major late and EIIa late upstream promoter regions do not contain such EIa-responsive sequence elements. In contrast, after transfection of these chimeric promoter recombinants into 293 cells (which constitutively express the EIa proteins), we found that their relative levels of transcription are similar and markedly different from those observed when they are cotransfected into HeLa cells with EIa protein-producing recombinants. We conclude that the efficiency of transcription from a given promoter in 293 cells is not necessarily related to the presence of a specific EIa-responsive element.

scholarly journals Participation of the yeast activator Abf1 in meiosis-specific expression of the HOP1 gene.

1996 ◽  
Vol 16 (6) ◽  
pp. 2777-2786 ◽  
Author(s):  
V Gailus-Durner ◽  
J Xie ◽  
C Chintamaneni ◽  
A K Vershon
Keyword(s):  
Transcriptional Activation ◽  
Mutational Analysis ◽  
Consensus Sequence ◽  
Promoter Sequence ◽  
Regulatory Elements ◽  
Specific Gene ◽  
Specific Expression ◽  
Autonomously Replicating Sequence

The meiosis-specific gene HOP1, which encodes a component of the synaptonemal complex, is controlled through two regulatory elements, UASH and URS1H. Sites similar to URS1H have been identified in the promoter region of virtually every early meiosis-specific gene, as well as in many promoters of nonmeiotic genes, and it has been shown that the proteins that bind to this site function to regulate meiotic and nonmeiotic transcription. Sites similar to the UASH site have been found in a number of meiotic and nonmeiotic genes as well. Since it has been shown that UASH functions as an activator site in vegetative haploid cells, it seemed likely that the factors binding to this site regulate both meiotic and nonmeiotic transcription. We purified the factor binding to the UASH element of the HOP1 promoter. Sequence analysis identified the protein as Abf1 (autonomously replicating sequence-binding factor 1), a multifunctional protein involved in DNA replication, silencing, and transcriptional regulation. We show by mutational analysis of the UASH site, that positions outside of the proposed UASH consensus sequence (TNTGN[A/T]GT) are required for DNA binding in vitro and transcriptional activation in vivo. A new UASH consensus sequence derived from this mutational analysis closely matches a consensus Abf1 binding site. We also show that an Abf1 site from a nonmeiotic gene can replace the function of the UASH site in the HOP1 promoter. Taken together, these results show that Abf1 functions to regulate meiotic gene expression.

scholarly journals Neuropeptide B mediates female sexual receptivity in medaka fish, acting in a female-specific but reversible manner

eLife ◽  
2019 ◽  
Vol 8 ◽  
Author(s):  
Towako Hiraki-Kajiyama ◽  
Junpei Yamashita ◽  
Keiko Yokoyama ◽  
Yukiko Kikuchi ◽  
Mikoto Nakajo ◽  
...  
Keyword(s):  
Mating Behavior ◽  
Transcriptional Activation ◽  
Sex Steroid ◽  
Critical Element ◽  
Specific Expression ◽  
Behavioral Studies ◽  
Estrogen Responsive Element ◽  
Mating Behaviors ◽  
Responsive Element ◽  
Female Specific

Male and female animals display innate sex-specific mating behaviors. In teleost fish, altering the adult sex steroid milieu can effectively reverse sex-typical mating behaviors, suggesting remarkable sexual lability of their brains as adults. In the teleost medaka, neuropeptide B (NPB) is expressed female-specifically in the brain nuclei implicated in mating behavior. Here, we demonstrate that NPB is a direct mediator of estrogen action on female mating behavior, acting in a female-specific but reversible manner. Analysis of regulatory mechanisms revealed that the female-specific expression of NPB is dependent on direct transcriptional activation by estrogen via an estrogen-responsive element and is reversed in response to changes in the adult sex steroid milieu. Behavioral studies of NPB knockouts revealed that female-specific NBP mediates female receptivity to male courtship. The female-specific NPB signaling identified herein is presumably a critical element of the neural circuitry underlying sexual dimorphism and lability of mating behaviors in teleosts.

scholarly journals Discordant Influence of Blastomyces dermatitidis Yeast-Phase-Specific Gene BYS1 on Morphogenesis and Virulence

2010 ◽  
Vol 78 (6) ◽  
pp. 2522-2528 ◽  
Author(s):  
Theerapong Krajaejun ◽  
Marcel Wüthrich ◽  
Gregory M. Gauthier ◽  
Thomas F. Warner ◽  
Thomas D. Sullivan ◽  
...  
Keyword(s):  
Signal Sequence ◽  
Yeast Cells ◽  
Blastomyces Dermatitidis ◽  
Mouse Lung ◽  
Specific Gene ◽  
Dimorphic Fungus ◽  
Yeast Phase ◽  
Different Strains

ABSTRACT Blastomyces dermatitidis is a thermally induced dimorphic fungus capable of causing lung and systemic infections in immunocompetent animal hosts. With the publication of genomic sequences from three different strains of B. dermatitidis and the development of RNA interference as a gene-silencing tool, it has become possible to easily ascertain the virulence and morphological effects of knocking down the expression of candidate genes of interest. BYS1 (Blastomyces yeast-phase-specific 1), first identified by Burg and Smith, is expressed at high levels in yeast cells and is undetectable in mold. The deduced protein sequence of BYS1 has a putative signal sequence at its N terminus, opening the possibility that the BYS1-encoded protein is associated with the yeast cell wall. Herein, strains of B. dermatitidis with silenced expression of BYS1 were engineered and tested for morphology and virulence. The silenced strains produced rough-surfaced cultures on agar medium and demonstrated a propensity to form pseudohyphal cells on prolonged culture in vitro and in vivo, as measured in the mouse lung. Tests using a mouse model of blastomycosis with either yeast or spore inocula showed that the bys1-silenced strains were as virulent as control strains. Thus, although silencing of BYS1 alters morphology at 37°C, it does not appear to impair the pathogenicity of B. dermatitidis.

scholarly journals Neuropeptide B mediates female sexual receptivity in medaka fish, acting in a female-specific but reversible manner

2019 ◽  
Author(s):  
Towako Hiraki-Kajiyama ◽  
Junpei Yamashita ◽  
Keiko Yokoyama ◽  
Kohei Hosono ◽  
Yukika Kawabata-Sakata ◽  
...  
Keyword(s):  
Mating Behavior ◽  
Transcriptional Activation ◽  
Sex Steroid ◽  
Critical Element ◽  
Specific Expression ◽  
Behavioral Studies ◽  
Estrogen Responsive Element ◽  
Mating Behaviors ◽  
Responsive Element ◽  
Female Specific

AbstractMale and female animals display innate sex-specific mating behaviors. Among vertebrates, teleosts are unique in that altering the adult sex steroid milieu can reverse sex-typical mating behaviors, suggesting sexual lability of their brains. In the teleost medaka, neuropeptide B (NPB) is expressed female-specifically in the brain nuclei implicated in mating behavior. Here, we demonstrate that NPB is a direct mediator of estrogen action on female mating behavior, acting in a female-specific but reversible manner. Analysis of regulatory mechanisms revealed that the female-specific expression of NPB is dependent on direct transcriptional activation by estrogen via an estrogen-responsive element and is reversed in response to changes in the adult sex steroid milieu. Behavioral studies of NPB knockouts revealed that female-specific NBP mediates female receptivity to male courtship. The female-specific NPB signaling identified herein is presumably a critical element of the neural circuitry underlying sexual dimorphism and lability of mating behaviors in teleosts.

scholarly journals Probing the Yeast Phase-Specific Expression of the CBP1 Gene in Histoplasma capsulatum

1998 ◽  
Vol 180 (7) ◽  
pp. 1786-1792 ◽  
Author(s):  
Jean Baldus Patel ◽  
Janet West Batanghari ◽  
William E. Goldman
Keyword(s):  
Calcium Binding ◽  
Histoplasma Capsulatum ◽  
Specific Activity ◽  
Pathogenic Fungus ◽  
Start Codon ◽  
Regulatory Sequences ◽  
Gene Encoding ◽  
Specific Expression ◽  
Promoter Sequences ◽  
Yeast Phase

ABSTRACT Histoplasma capsulatum is a pathogenic fungus that exists in two distinct forms. The saprophytic mycelial phase inhabits moist soil environments; once inhaled, hyphae and conidia convert to a unicellular yeast phase that is capable of parasitizing macrophage phagolysosomes. Yeasts cultures, but not mycelial cultures, release large quantities of a calcium-binding protein (CBP) which may be important in calcium acquisition during intracellular parasitism. In this study, we show that the gene encoding CBP (CBP1) is transcriptionally regulated. To identify promoter sequences that are important for yeast phase-specific activity, we created a series of fusions between successively truncatedCBP1 5′ untranslated regulatory sequences and theEschericha coli lacZ gene. The fusions were constructed on a telomeric shuttle plasmid that can replicate autonomously in the fungus. By assaying for β-galactosidase activity from H. capsulatum transformants, we identified a 102-bp region that mediates promoter activation and yeast phase promoter activity. Base pair substitution analysis suggests that the sequences between 839 and 877 bp upstream of the start codon are the most important for this positive regulation.

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