(Theme 5) You are testing the role of three proteins (Protein A, Protein B, and Protein C) which have potential roles in transcriptional regulation of a gene in a mammalian cell line. You do this using a gel shift assay. Here, purified proteins are mixed in a test tube with a labeled DNA fragment that contains a putative regulatory region found upstream of a protein-coding gene. Decrea in the mobility of the DNA (ie it cannot move as far through the gel from top to bottom as shown) can be inferred to reflect the effect of stable DNA-protein binding interactions. You perform s reactions, with the added proteins indicated above the gel (+ = protein present; - = protein absent). Low DNA Mobility High 1 2 3 4 + 5 + + 6 Protein A Protein B Protein C Lane a) From the data shown which protein(s) directly bind the DNA? Briefly justify your answer. b) Considering your answer in A, briefly explain the results in lanes 5 and 6. c) Additional genetic data indicates that two of these three proteins are both required to activate transcription of the gene. Briefly describe which two proteins this would be, and propose a model for the specific molecular role of each.

(Theme 5) You are testing the role of three proteins (Protein A, Protein B, and Protein C) which have potential roles in transcriptional regulation of a gene in a mammalian cell line. You do this using a gel shift assay. Here, purified proteins are mixed in a test tube with a labeled DNA fragment that contains a putative regulatory region found upstream of a protein-coding gene. Decrea in the mobility of the DNA (ie it cannot move as far through the gel from top to bottom as shown) can be inferred to reflect the effect of stable DNA-protein binding interactions. You perform s reactions, with the added proteins indicated above the gel (+ = protein present; - = protein absent). Low DNA Mobility High 1 2 3 4 + 5 + + 6 Protein A Protein B Protein C Lane a) From the data shown which protein(s) directly bind the DNA? Briefly justify your answer. b) Considering your answer in A, briefly explain the results in lanes 5 and 6. c) Additional genetic data indicates that two of these three proteins are both required to activate transcription of the gene. Briefly describe which two proteins this would be, and propose a model for the specific molecular role of each.

Human Heredity: Principles and Issues (MindTap Course List)

11th Edition

ISBN:9781305251052

Author:Michael Cummings

Publisher:Michael Cummings

Chapter9: Gene Expression And Gene Regulation

Section: Chapter Questions

Problem 10QP: The pre-mRNA transcript and protein made by several mutant genes were examined. The results are...

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