Cellular localization of steroid hormone-regulated proteins during sexual development in Achlya

Brunt, Shelley A.; Silver, Julie C.

doi:10.1016/0014-4827(86)90585-9

Cited by 31 publications

(31 citation statements)

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“…Cells of A. ambisexualis J. Raper strain E87 were grown as described previously (49 Hormone treatment and in vivo labeling conditions. Cells were grown as described previously (9)(10)(11) (47). Cell homogenates were filtered through Miracloth, and the crude filtrate was treated as described by Brunt and Silver (10) to obtain the cytoplasmic and crude nuclear pellet.…”

Section: Materuils and Methodsmentioning

confidence: 99%

“…Cell homogenates were filtered through Miracloth, and the crude filtrate was treated as described by Brunt and Silver (10) to obtain the cytoplasmic and crude nuclear pellet. Total nuclear proteins were prepared from the further-purified nuclear fraction as previously described (10,49).…”

Section: Materuils and Methodsmentioning

confidence: 99%

“…changes in the synthesis or processing of certain Achlyasecreted glycoproteins (9). One of the prominent antheridiolinduced proteins exhibits a molecular weight of 85,000 and was found both in the nuclear and cytoplasmic fractions of antheridiol-treated cells (10). This protein seemed very similar in electrophoretic behavior to the Achlya 85-kilodalton (kDa) heat shock protein (49), which is one of the several characteristic heat shock proteins observed in A. ambisexualis (33,34,49).…”

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confidence: 95%

“…Studies in our laboratory have focused on the early developmental events which occur when antheridiol is added to vegetatively growing cultures of the male mating type, strain E87 (9)(10)(11)48). In E87, one of the earliest developmental events seen at the morphological level after treatment of cells with the hormone is the formation of distinct structures called antheridial branches (3).…”

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confidence: 99%

“…These proteins could be localized to specific cell fractions (9)(10)(11). The hormone also appears to induce * Corresponding author.…”

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Steroid hormone regulation of the Achlya ambisexualis 85-kilodalton heat shock protein, a component of the Achlya steroid receptor complex.

Brunt¹,

Riehl²,

Silver³

1990

Mol. Cell. Biol.

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The steroid hormone antheridiol regulates sexual development in the fungus Achlya ambisexualis. Analyses of in vivo-labeled proteins from hormone-treated cells revealed that one of the characteristic antheridiolinduced proteins appeared to be very similar to the Achlya 85-kilodalton (kDa) heat shock protein. Analysis of in vitro translation products of RNA isolated from control, heat-shocked, or hormone-treated cells demonstrated an increased accumulation of mRNA encoding a similar 85-kDa protein in both the heat-shocked and hormone-treated cells. Northern (RNA) blot analyses with a Drosophila melanogaster hsp83 probe indicated that a mRNA species of approximately 2.8 kilobases was substantially enriched in both heat-shocked and hormone-treated cells. The monoclonal antibody AC88, which recognizes the non-hormone-binding component of the Achlya steroid receptor, cross-reacted with Achlya hsp85 in cytosols from heat-shocked cells. This monoclonal antibody also recognized both the hormone-induced and heat shock-induced 85-kDa in vitro translation products. Taken together, these data suggest that similar or identical 85-kDa proteins are independently regulated by the steroid hormone antheridiol and by heat shock and that this protein is part of the Achlya steroid receptor complex. Our results demonstrate that the association of hsp9O family proteins with steroid receptors observed in mammals and birds extends also to the eucaryotic microbes and suggest that this association may have evolved early in steroid-responsive systems.The water mold Achlya ambisexualis is a eucaryotic, filamentous fungus in which sexual reproduction is regulated by well-characterized steroid hormones (4,22,54). There are at least two mating types in A. ambisexualis, and these are usually referred to as male and female. Each of these mating types produces a specific fungal steroid hormone which is secreted into the medium and which mediates both chemotropic and developmental changes in cells of the opposite mating type. In this respect, the Achlya steroids act as both pheromones and hormones.The fungal steroid hormone antheridiol, normally produced by female strains of A. ambisexualis, has been isolated (30) and characterized (2). Studies in our laboratory have focused on the early developmental events which occur when antheridiol is added to vegetatively growing cultures of the male mating type, strain E87 (9-11, 48). In E87, one of the earliest developmental events seen at the morphological level after treatment of cells with the hormone is the formation of distinct structures called antheridial branches (3). Both the time of onset of branching and the percent of hyphae branched are directly related to the amount of hormone added (9, 30). Thus, the response is very synchronous, and a high percentage of the hyphae is target tissue for the hormone. For these reasons, A. ambisexualis represents an interesting system in which to investigate both the mechanism of action of steroid hormones and the evolution of steroid hormone systems.In previou...

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Section: Materuils and Methodsmentioning

confidence: 99%

Section: Materuils and Methodsmentioning

confidence: 99%

mentioning

confidence: 95%

mentioning

confidence: 99%

“…These proteins could be localized to specific cell fractions (9)(10)(11). The hormone also appears to induce * Corresponding author.…”

mentioning

confidence: 99%

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Steroid hormone regulation of the Achlya ambisexualis 85-kilodalton heat shock protein, a component of the Achlya steroid receptor complex.

Brunt¹,

Riehl²,

Silver³

1990

Mol. Cell. Biol.

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Regulation of two different hsp70 transcript populations in steroid hormone‐induced fungal development

et al. 1993

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In the filamentous oomycete fungus Achlya, the differentiation of gamete bearing structures on vegetative hyphae of the male mating type, is induced by the Achlya steroid hormone, antheridiol. Among the several metabolically labeled intracellular proteins whose synthesis or accumulation is altered by hormone treatment are steroid-induced 85-kDa and 68- to 78-kDa proteins. The 85-kDa protein was previously shown to be the Achlya heat shock protein hsp85 [Brunt et al., 1990; Brunt and Silver, 1991], a component of the putative Achlya steroid hormone receptor. It was of interest to determine if the antheridiol-induced "70-kDa" proteins were hsp70-family heat shock proteins and if hormone treatment-induced changes in the level of hsp70 transcripts. Two different Achlya hsp70 genomic sequences were cloned and used to investigate these questions. The two hsp70 sequences recognized two different mycelial transcript populations, one of which was regulated also by decreased glucose. Of note, both of the two hsp70 transcript populations were found to be regulated by antheridiol. The hormone-induced changes in hsp70 transcript levels were temporally correlated with the onset of massive lateral hyphal branching and alterations in the pattern of secreted N-linked glycoproteins which occur in hormone-treated mycelia. To our knowledge, this represents one of the first reports on changes in hsp70 proteins and transcripts during fungal differentiation. Our results may have implications for the role of heat shock proteins in hyphal branching and secretion in filamentous fungi and perhaps other cell types.

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Regulation of hsp90 and hsp70 Genes during Antheridiol-Induced Hyphal Branching in the OomyceteAchlya ambisexualis

Brunt

Borkar

Silver

1998

Fungal Genetics and Biology

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Cellular localization of steroid hormone-regulated proteins during sexual development in Achlya

Cited by 31 publications

References 34 publications

Steroid hormone regulation of the Achlya ambisexualis 85-kilodalton heat shock protein, a component of the Achlya steroid receptor complex.

Steroid hormone regulation of the Achlya ambisexualis 85-kilodalton heat shock protein, a component of the Achlya steroid receptor complex.

Regulation of two different hsp70 transcript populations in steroid hormone‐induced fungal development

Regulation of hsp90 and hsp70 Genes during Antheridiol-Induced Hyphal Branching in the OomyceteAchlya ambisexualis

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