16 Aug Question: The C-terminal Domain (CTD) Of The Large Subunit Of Yeast RNA Polymerase II (RPB1) With Its Repeat-structures Is Important For Polymerase Function. A Protein (called Rtt103), Which Binds To A Phosphorylated Peptide With The Sequence YpSPTSPS From The CTD, But Not To Nonphosphorylated YSPTSPS, Was Isolated From Yeast Extracts And Identified By Mass Spectroscopy….
The C-terminal Domain (CTD) of the large subunit of yeast RNA polymerase II (RPB1) with its repeat-structures is important for polymerase function. A protein (called Rtt103), which binds to a phosphorylated peptide with the sequence YpSPTSPS from the CTD, but not to nonphosphorylated YSPTSPS, was isolated from yeast extracts and identified by mass spectroscopy. An epitope-tagged version of Rtt103 was used to find interacting proteins, and two such proteins, Rat1 and Rai1, were found. Question 5-1. Describe briefly how you would find interacting proteins. Question 5-2. Draw a sketch of the Rtt103 complex. From previously published investigations, it is known that Rat1 has a 5’->3’ RNA exonuclease activity, and Rai1 stimulates the activity of Rat1 in vitro. To investigate the function of Rtt103, Rat1 and Rai1, a number of Chromatin ImmunoPrecipitation (ChIP) analyses were carried out where cross-linked, sonicated chromatin is immunoprecipitated with a specific antibody and washed, original cross-links broken, and the presence of specific sequences investigated by PCR analysis. Several genes were investigated by ChIP using antibodies against Rtt103, Rat1 and Rai1. One of these was the EKS gene, and the positions of nine sequences on the EKS gene investigated are indicated in figure 5-1. The EKS-gene box represents all gene sequences, from the beginning of the first to the end of the last exon, including intronsThe C-terminal Domain (CTD) of the large subunit of yeast RNA polymerase II (RPB1) with its repeat-structures is important for polymerase function. A protein (called Rtt103), which binds to a phosphorylated peptide with the sequence YpSPTSPS from the CTD, but not to nonphosphorylated YSPTSPS, was isolated from yeast extracts and identified by mass spectroscopy. An epitope-tagged version of Rtt103 was used to find interacting proteins, and two such proteins, Rat1 and Rai1, were found.
Question 5-1. Describe briefly how you would find interacting proteins.
Question 5-2. Draw a sketch of the Rtt103 complex.
From previously published investigations, it is known that Rat1 has a 5’->3’ RNA exonuclease activity, and Rai1 stimulates the activity of Rat1 in vitro. To investigate the function of Rtt103, Rat1 and Rai1, a number of Chromatin ImmunoPrecipitation (ChIP) analyses were carried out where cross-linked, sonicated chromatin is immunoprecipitated with a specific antibody and washed, original cross-links broken, and the presence of specific sequences investigated by PCR analysis. Several genes were investigated by ChIP using antibodies against Rtt103, Rat1 and Rai1. One of these was the EKS gene, and the positions of nine sequences on the EKS gene investigated are indicated in figure 5-1. The EKS-gene box represents all gene sequences, from the beginning of the first to the end of the last exon, including introns
Question 5-3. What do the primers from the non-transcribed region in the upper panel control for?
Question 5-4. What does the experiments show about the proteins Rtt103, Rat1 and Rai1?
Our website has a team of professional writers who can help you write any of your homework. They will write your papers from scratch. We also have a team of editors just to make sure all papers are of HIGH QUALITY & PLAGIARISM FREE. To make an Order you only need to click Ask A Question and we will direct you to our Order Page at WriteDemy. Then fill Our Order Form with all your assignment instructions. Select your deadline and pay for your paper. You will get it few hours before your set deadline.
Fill in all the assignment paper details that are required in the order form with the standard information being the page count, deadline, academic level and type of paper. It is advisable to have this information at hand so that you can quickly fill in the necessary information needed in the form for the essay writer to be immediately assigned to your writing project. Make payment for the custom essay order to enable us to assign a suitable writer to your order. Payments are made through Paypal on a secured billing page. Finally, sit back and relax.
About Wridemy
We are a professional paper writing website. If you have searched a question and bumped into our website just know you are in the right place to get help in your coursework. We offer HIGH QUALITY & PLAGIARISM FREE Papers.
How It Works
To make an Order you only need to click on “Order Now” and we will direct you to our Order Page. Fill Our Order Form with all your assignment instructions. Select your deadline and pay for your paper. You will get it few hours before your set deadline.
Are there Discounts?
All new clients are eligible for 20% off in their first Order. Our payment method is safe and secure.