Rapidly labeled proteins on the salivary gland chromosomes of Drosophila melanogaster

Mitchell, Herschel K.; Lipps, Loveriza S.

doi:10.1007/bf00484917

Cited by 45 publications

(14 citation statements)

References 12 publications

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“…Furthermore, evidence of control of protein synthesis at the transcriptional level was also presented. By pulse-labeling of salivary gland proteins, Mitchell and Lipps [9] confirmed the conclusion of Berendes [3] that protein accumulation precedes RNA synthesis at a puff site. Specifically, at 0 h prepupae pulse-labeled salivary gland showed an accumulation of label at 85D just about an hour before it is due to puff.…”

Section: Introductionmentioning

confidence: 68%

Drosophila salivary gland proteins and pupation

Sarmiento

Mitchell

1982

Dev. Genet.

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Pulse-labeling experiments of salivary glands from the prepupal stages of development showed selectively high rates of synthesis of a set of low molecular weight proteins (6K-12K). These proteins are stably maintained in the salivary glands during prepupal development and are subsequently transported to the pupation fluid (found between the pupal case and the prepupal cuticle) when pupation occurs. These small polypeptides are very basic with the major components having isoelectric points of 8.6-8.7 and the minor components having isoelectric points of 9.1-9.5. This study shows the continuing function of the salivary glands-specifically, the synthesis and secretion of a set of proteins with a putative role in pupation.

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Section: Introductionmentioning

confidence: 68%

Drosophila salivary gland proteins and pupation

Sarmiento

Mitchell

1982

Dev. Genet.

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“…Previous reports have indicated that Drosophila heat-shock proteins are transported into the nucleus soon after synthesis (16,31). Vincent and Tanguay (32) have also shown in Chironomous tentans by microdissection that some heatshock proteins exhibit a broad intranuclear distribution.…”

Section: Discussionmentioning

confidence: 92%

Association between the mammalian 110,000-dalton heat-shock protein and nucleoli.

Subjeck¹,

Shyy²,

Jing³

et al. 1983

The Journal of Cell Biology

112

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A rabbit antiserum has been prepared using as antigen the 110,000-dalton mammalian heat-shock protein. This protein was purified for injection by two-dimensional PAGE of heat-shocked Chinese hamster ovary cells. Characterization by immunoautoradiography and immunoprecipitation reveals that the antiserum is specific for the 110,000-dalton protein.Both techniques also reveal that the protein against which the antiserum is directed is induced by heat shock. Indirect immunofluorescence shows that the antigen is primarily localized at or near the nucleolus in cultured cells and numerous murine tissues. Treatment of cultured cells with deoxyribonuclease destroys the organization of staining within the nucleus while ribonuclease appears to completely release the antigen from the nucleus. A binding of the antiserum to cytoplasmic structures is also observed by immunofluorescence. This association with nucleoli may have implications in the regulatory aspects of the heat-shock response.A hyperthermic challenge to a cell can result in a reorganization at translational and transcriptional levels (7, 23) and at the same time protect the cell from additional applications of the same stress (8), probably through the intervention of heatshock proteins (14,15,17,18,27,28). Despite the implications of this, it is important to recognize that the function of heat-shock proteins remains a mystery. However, reports indicate that at least some heat-shock proteins my localize in the nucleus (2,3,13,16,31,32), suggesting a direct role in the regulatory changes associated with this response.To learn more regarding the function of heat-shock proteins and their role in regulation and protection, we investigated mammalian heat-shock proteins using an immunologic approach. We report here on the characterization of an antiserum against the major l l0,000-dalton mammalian heatshock protein (HSP 110). ~ It is demonstrated by indirect immunofluorescence that this protein is localized at or near the nucleolus and is released from the nucleus by treatment with ribonuclease. This suggests that aspects of nucleolar function may be important in the protective and regulatory properties of the heat-shock response. mented with 15% heat-inactivated newborn calf serum. In heat-shock experiments: flasks were immersed horizontally into a constant temperature water bath (Haake FK-2) for the indicated times at 45"C + 0.1°C. 10TY2 mouse embryo fibroblasts were originally obtained from Dr. John Bertram of this Institute. These cells are grown in Eagles basal medium with Earle's salts (Gibco Laboratories) supplemented with 10% heat-inactivated fetal calf serum. Frozen sections were obtained from tissues of BALB/c CR mice supplied by the West Seneca Laboratories of this Institute. MATERIALS AND METHODS Cells Preparation of Antigen and Immunoautoradiography:Heat-shock proteins were identified by autoradiography of two-dimensional O'Farrel gels, as previously modified and described (10,19), by applying a [35S]methionine pulse during the peak induction peri...

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“…Samples for observing bristle morphology were prepared as described by Mitchell and Lipps [l] , and those for observing chromosome autoradiographs were made as described earlier [ 1,9] .…”

Section: Scanning Electron Microscopementioning

confidence: 99%

“…These various facts show different kinds of correlations between the presence of heat-shock proteins and protection phenomena. They suggest but do not prove that the heat-shock protein(s) is directly involved physically with the storage of mRNA and/or the regulation of protein synthesis just as they have been shown to be involved directly with salivary gland chromosomes [9]. There now seems little doubt that heat shock does affect storage of the message and preferential translation of the message after heat shock is well established [12,13,141.…”

Section: Protein Synthesis and Protection For Survivalmentioning

confidence: 99%

“…The evidence presented here supports both the existence of a cytoplasmic pool of RNPs and the postulate that such RNPs are fundamental importance in the regulation of development. We have added the suggestion of a direct involvement of heat-shock protein(s) in the system both in the nucleus [9] and in the cytoplasm, but this may be only part of the normal picture. It is of interest also to note that the heat-shock phenomenon (the induction of synthesis of specific proteins) has now been observed in a variety of cell-types other than Drosophila [23,26] .…”

Section: A Speculative Modelmentioning

confidence: 99%

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Specific protection from phenocopy induction by heat shock

et al. 1979

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Mild heat treatments applied to whole animals or cell cultures of Drosophila prior to lethal heat shocks result in increased survival and protection against phenocopy induction. The optimal condition for the preliminary mild heat treatment is that which induces the synthesis of heat-shock proteins but does not turn off the protein synthesis that is in progress. Recovery of protein synthesis but not RNA synthesis following a drastic heat shock is much enhanced by the pretreatments. The results suggest that the protection for survival and against phenocopy induction is due to storage of messenger RNA.Key words: drosophila, gene regulation, heat shock, protection phenocopies, survival I NTROD UCTl ON We recently described [ I ] a series of phenocopies that are induced in Drosophila melanogaster by subjecting pupa to heat shock at specific stages of development. Three of the phenocopies produced by shocks at successive intervals closely resemble the mutants hook and javelin which are involved in determining the structure of the scutellar bristles of the adult fly. We have also shown that the conditions used for phenocopy induction, turn off, for a time, both transciptional and translational activities in Drosophila tissues [ 1 , 2 ] . Subsequently, translation resumes long before reactivation of transcription [ l ] . These findings show that heat shock can result in storage of mRNAs in keeping with the observations of McKenzie et a1 [ 131 and Mirault et a1 [14] . Furthermore these observations provide a reasonable basis for interpretation of the heat-shock protection phenomenon described by Milkman several years ago [3,4] . Milkman and collaborators demonstrated that when Drosophila pupa (24 hours) were subjected to a mild heat shock prior to one that would be lethal alone, some animals would survive. They also reported that a pretreatment of this kind would prevent the induction of a phenocopy of the mutant crossveinless.

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Rapidly labeled proteins on the salivary gland chromosomes of Drosophila melanogaster

Cited by 45 publications

References 12 publications

Drosophila salivary gland proteins and pupation

Drosophila salivary gland proteins and pupation

Association between the mammalian 110,000-dalton heat-shock protein and nucleoli.

Specific protection from phenocopy induction by heat shock

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