Visualizing Cellular Stress: A Hypothesis-Driven Confocal Laboratory Exercise to Identify Compounds That Activate Heat Shock Factor Binding at "Hsp70" Loci

Autor/inn/en	Choi, Annette; Wang, Mengqi; Hrizo, Stacy; Buckley, Martin S.
Titel	Visualizing Cellular Stress: A Hypothesis-Driven Confocal Laboratory Exercise to Identify Compounds That Activate Heat Shock Factor Binding at "Hsp70" Loci
Quelle	In: Biochemistry and Molecular Biology Education, 46 (2018) 5, S.445-452 (8 Seiten)Infoseite zur Zeitschrift PDF als Volltext Verfügbarkeit
Zusatzinformation	ORCID (Buckley, Martin S.)
Sprache	englisch
Dokumenttyp	gedruckt; online; Zeitschriftenaufsatz
ISSN	1470-8175
DOI	10.1002/bmb.21163
Schlagwörter	Leitfaden; Unterricht; Lehrer; Heat; Science Laboratories; Genetics; Cytology; Molecular Biology; Hypothesis Testing; Entomology; Science Instruction; Laboratory Equipment; Laboratory Experiments; College Science; College Students + Suchen Sie Ihr Suchwort? Lesson concept; Instruction; Unterrichtsentwurf; Unterrichtsprozess; Teacher; Teachers; Lehrerin; Lehrende; Hitze; Humangenetik; Zytologie; Molekularbiologie; Hypothesenprüfung; Hypothesentest; Entomologie; Teaching of science; Science education; Natural sciences Lessons; Naturwissenschaftlicher Unterricht; Laborausstattung; Laboratory work; Laborarbeit; Collegestudent
Abstract	Exposure of organisms to high temperatures and various chemical and physical stressors can cause protein misfolding and aggregation. In turn, this can disrupt the functions of proteins, threatening both development and homeostasis. To overcome this, cells can initiate the highly conserved heat shock (HS) stress response pathway. In eukaryotes, this is a coordinated cellular response, in which the master HS activator, heat shock factor (HSF), is rapidly recruited to the HS protein genes, and triggers the recruitment of additional coactivator proteins that facilitate gene expression. This results in the production of HS proteins that function as nuclear and cytosolic molecular chaperones, to promote refolding of proteins and prevent aggregation and increase protein degradation pathways. Here, we describe a laboratory exercise in which students visualize and quantify Green Fluorescent Protein (GFP)-tagged HSF binding to the HS protein genes in living "Drosophila" salivary gland nuclei as an output of chemically induced protein misfolding. Students are assigned an array of chemicals, and using the scientific literature, predict impacts of these chemicals on protein folding. Students then test the effects of their chemicals by measuring GFP-tagged HSF binding to the HS genes in salivary glands using confocal microscopy. Designed for junior and senior level students in a cell/molecular biology course, this is a two-part lab, in which student work closely with an instructor to help familiarize them with developing hypotheses supported by scientific literature and testing these hypotheses by quantitating the levels of GFP-HSF binding, using confocal microscopy of living "Drosophila" cells. (As Provided).
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Erfasst von	ERIC (Education Resources Information Center), Washington, DC
Update	2020/1/01