NPH1 - B) Move nph1 expression from the membrane to the...

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“NPH” = nonphototropic hypocotyl NPH1 ATP Cell membrane outside inside NPH1 ADP How do receptors work? Blue light receptor. NPH1 absorbs blue light Triggers phosphorylation ATP -> ADP …does not require a protein kinase.
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How did they figure out the role of NPH1? (Briggs’ lab, Carnegie Inst.) Step 1 : Discover a membrane protein that is phosphorylated (acquires a PO 4 2- from ATP) in response to blue light. Step 2 : Screen for Arabidopsis mutants that do not bend toward light. • Found 4 different mutant types. • One type (nph1) does not have the membrane protein (from Step 1). Wild type nph mutant
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iClicker experimental design: If you wanted to prove that nph1 autophosphorylates in the presence of blue light, what experiment would you do? A) Knock out nph1 and expose the plant to blue light.
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Unformatted text preview: B) Move nph1 expression from the membrane to the vacuole. C) Clone functional nph1 into an nph1 knockout mutant. D) Insert the nph1 gene into insect cells. How did they figure out the role of NPH1? (Briggs’ lab, Carnegie Inst.) Step 1 : Discover a membrane protein that is phosphorylated (acquires a PO 4 2-from ATP) in response to blue light. Step 2 : Screen for Arabidopsis mutants that do not bend toward light. • Found 4 different mutant types. • One type (nph1) does not have the membrane protein (from Step 1). Wild type nph mutant Step 3 : Insert nph1 wild-type gene into insect(!) cells • NPH1 protein, which is phosphorylated in blue light. • Does not require other catalysts = “autophosphorylation”...
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This note was uploaded on 04/06/2008 for the course BIO 20b taught by Professor Dunkin&parker during the Winter '08 term at University of California, Santa Cruz.

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NPH1 - B) Move nph1 expression from the membrane to the...

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