NE102 Lecture Notes 2

Many transcription factors interact with hats or

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Many transcription factors interact with HATs or HDACs
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HATs recruited by transcriptional activators -> chromatin de-condensation ->  transcriptional activation HATs = “Co=activators” HDACs recruited by transcriptional repressors -> chromatin condensation ->  transcriptional repression HDACs = “Co-repressors” Histone modifying enzymes also shown to bind to the RNA pol II CTD SPECIFIC NEURO EXAMPLE? Regulation of gene expression by the transcription factor CREB plays a critical role  during long-term potentiation CREB regulates numerous genes involved in neurotransmission CREB is always in the nucleus  bound to its transcription site (Inactive or not) CREB activates transcription of its target genes by recruiting the co-activator  CREB- binding protein Protein-protein interactions between CREB & CBP require phosphorylation of CREB Phosphorylated by several kinases, including  Ca+2/calmodulin kinase IV (CaMKIV) Opening of NMDA leads to flooding of Calcium ions that activates the kinase CaMKIV
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Antibodies as Probes for Proteins 19:00 QUESTION: How can we visualize & study specific proteins in cells? Anitbodies  can be used as “probes”.  PREREQUISITE QUESTION: What are “anitbodies”? Proteins produced by the immune system Bind to foreign substances (i.e., antigens) which targets them for attack by immune  system cells. The immune system can produce millions of different antibodies that can bind millions of  different antigens Y-shaped structure Have two antigen binding sites at the tips Epitope : the spot where the antibody binds to the antigen Constant region : Part of antibody that is the same for all antibodies Variable region : Part of the antibody that differs from antibody to antibody Difference is what dictates which specific antigen the antibody will bind. Antibodies that bind a desire antigen can be generated by inoculating an animal (rabbit,  mouse, or other) with that antigen The antibodies can then be purified from the animal and used as probes for your protein  of interest. Once purified, antibodies can be used to visualize your protein of interest. QUESTION: How? Once purified, antibodies can be conjugated onto molecules that  emit light . (e.g.,  fluorophores that emit fluorescence) Fluorophore-conjugated anitbodies can be used to examine WHERE your proteins of  interest is localize in cells via  immunocytochemsity
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Antibodies as Probes for Proteins 19:00 IMMUNOCYTOCHEMISTRY Four steps: 1.  Cells are “fixed” with a chemical “cross-linker” (e.g., formaldehyde) a. Killing the cells and fixing the proteins in their place.
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