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Questions and Answers
What is the purpose of the C-terminal tag in the Rrp44 expression plasmids?
What is the purpose of the C-terminal tag in the Rrp44 expression plasmids?
Which buffer was used for exosome purification during the experiments?
Which buffer was used for exosome purification during the experiments?
What method was used to create point mutations in the Rrp44 expression plasmids?
What method was used to create point mutations in the Rrp44 expression plasmids?
What was the method used to normalize enzyme concentrations during the experiments?
What was the method used to normalize enzyme concentrations during the experiments?
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What effect does treating the samples with 800 mM MgCl2 have during the purification process?
What effect does treating the samples with 800 mM MgCl2 have during the purification process?
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What is the dual function of the C-terminal S1 RNA-binding domain?
What is the dual function of the C-terminal S1 RNA-binding domain?
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What is the consequence of a mutation like G916E on the exosome complex?
What is the consequence of a mutation like G916E on the exosome complex?
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Which of the following statements is true about Rrp44?
Which of the following statements is true about Rrp44?
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What happens if a single core component of the exosome is depleted?
What happens if a single core component of the exosome is depleted?
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What is the importance of the exosome complex?
What is the importance of the exosome complex?
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Which technique was used to analyze the growth of yeast strains expressing Rrp44?
Which technique was used to analyze the growth of yeast strains expressing Rrp44?
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What does the presence of the GAL::rrp44 strain suggest about Rrp44's role?
What does the presence of the GAL::rrp44 strain suggest about Rrp44's role?
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What was utilized for the visualization of specific RNA sequences in the study?
What was utilized for the visualization of specific RNA sequences in the study?
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What was the outcome of transferring the GAL::rrp44 strain to glucose medium?
What was the outcome of transferring the GAL::rrp44 strain to glucose medium?
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What effect does deletion of the RRP44 ORF have on its function?
What effect does deletion of the RRP44 ORF have on its function?
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How did the doubling time of the rrp44D strains compare when complemented by Rrp44-exo and WT Rrp44?
How did the doubling time of the rrp44D strains compare when complemented by Rrp44-exo and WT Rrp44?
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What characteristic defect is associated with genetic depletion of Rrp44?
What characteristic defect is associated with genetic depletion of Rrp44?
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What was indicated about Rrp44's exonuclease activity based on the study?
What was indicated about Rrp44's exonuclease activity based on the study?
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Which plasmid resistance marker was used when constructing the rrp44D strain?
Which plasmid resistance marker was used when constructing the rrp44D strain?
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What happens to cells expressing Ura3 when they lose the URA3 plasmid?
What happens to cells expressing Ura3 when they lose the URA3 plasmid?
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What significant result was observed when comparing strains expressing only Rrp44-exo to those expressing WT Rrp44?
What significant result was observed when comparing strains expressing only Rrp44-exo to those expressing WT Rrp44?
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What mutation is associated with Rrp44 that allows it to generate degradation intermediates?
What mutation is associated with Rrp44 that allows it to generate degradation intermediates?
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What is suggested about the degradation activity of Rrp44-exo in the presence of Mn2+ ions?
What is suggested about the degradation activity of Rrp44-exo in the presence of Mn2+ ions?
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What aspect differentiates Ssd1-v from Ssd1-d in yeast strains?
What aspect differentiates Ssd1-v from Ssd1-d in yeast strains?
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What does the Rrp44-exo mutation paired with point mutations at conserved active-site amino acids create?
What does the Rrp44-exo mutation paired with point mutations at conserved active-site amino acids create?
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What effect did the depletion of intact Rrp44 have on the growth of W303 strains?
What effect did the depletion of intact Rrp44 have on the growth of W303 strains?
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How was RNA degradation monitored in the study?
How was RNA degradation monitored in the study?
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What is the role of the PIN domain in the context of Rrp44 activity?
What is the role of the PIN domain in the context of Rrp44 activity?
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What does the genetic interaction of Ssd1 indicate about its function?
What does the genetic interaction of Ssd1 indicate about its function?
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What role do the alterations in the C-terminal regions of Rrp44 play concerning the exosome?
What role do the alterations in the C-terminal regions of Rrp44 play concerning the exosome?
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Which of the following conclusions can be drawn about Rrp6 and Rrp44 interactions?
Which of the following conclusions can be drawn about Rrp6 and Rrp44 interactions?
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What distinguishes the functions of eukaryotic Rrp44 from bacterial RNase R?
What distinguishes the functions of eukaryotic Rrp44 from bacterial RNase R?
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Which of the following is a characteristic of the eukaryotic exosome?
Which of the following is a characteristic of the eukaryotic exosome?
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What activity does the PIN domain of Rrp44 appear to link?
What activity does the PIN domain of Rrp44 appear to link?
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In E. coli, what does PNPase associate with in the degradosome complex?
In E. coli, what does PNPase associate with in the degradosome complex?
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What influence does the balance between RNA processing and decay have?
What influence does the balance between RNA processing and decay have?
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What significant role do the proteins of the Rex family play within the exosome context?
What significant role do the proteins of the Rex family play within the exosome context?
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What role does the SKI2 DEVH box protein have in yeast?
What role does the SKI2 DEVH box protein have in yeast?
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Which element is associated with the stable maintenance of chromosomes in yeast?
Which element is associated with the stable maintenance of chromosomes in yeast?
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What is the primary use of the versatile toolbox for PCR-based tagging in yeast genes?
What is the primary use of the versatile toolbox for PCR-based tagging in yeast genes?
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Which protein complex is linked to pre-mRNA splicing and identified as a general splicing mutant suppressor?
Which protein complex is linked to pre-mRNA splicing and identified as a general splicing mutant suppressor?
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What type of activity is enhanced by Dis3 in relation to RCC1?
What type of activity is enhanced by Dis3 in relation to RCC1?
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Which process does the 30 to 50 exonucleases involve in yeast according to the context provided?
Which process does the 30 to 50 exonucleases involve in yeast according to the context provided?
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What is the function of the additional modules mentioned for PCR-based gene manipulation?
What is the function of the additional modules mentioned for PCR-based gene manipulation?
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What is one role associated with the RNA binding protein Ssd1p in yeast?
What is one role associated with the RNA binding protein Ssd1p in yeast?
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Study Notes
Yeast Core Exosome and Rrp44
- Yeast exosome components are shared between nuclear and cytoplasmic forms.
- Rrp44 (Dis3) is a key 3'-5' exonuclease in the yeast core exosome.
- Rrp44 lacking exonuclease activity (Rrp44-exo) supports growth.
- Rrp44-exo is synthetic-lethal with loss of cytoplasmic 5'-exonuclease Xrn1, but not with loss of nuclear 3'-exonuclease Rrp6.
- Rrp44-exo has a milder RNA processing phenotype than Rrp44 depletion.
- Recombinant Rrp44 has manganese-dependent endonuclease activity.
- The endonuclease activity is located in the N-terminal PIN domain.
- Loss of exonuclease activity does not immediately cause synthetic lethality.
- Loss of exonuclease activity of Rrp44 and Rrp6 results in reduced growth
- Rrp44 is crucial for pre-rRNA maturation.
- The exonuclease activity of Rrp44 is less critical for early steps in pre-rRNA processing.
Rrp44 Endonuclease Activity
- Rrp44 has both exonuclease and endonuclease activity, depending on Mn2+ presence.
- The N-terminal PIN domain is necessary and sufficient for exosome association.
- Four conserved amino acids within the PIN domain bind Mn2+ and are essential for endonuclease activity.
- The exonuclease activity is located in the RNB domain.
- Rrp44-exo mutation shows no strong lethality with loss of Rrp6.
- But Rrp44-exo is synthetically lethal with cytoplasmic 5'-exonuclease Xrn1.
Structural and Functional Role of Rrp44
- The PIN domain of Rrp44 is required for exosome association.
- The S1 RNA-binding domain is important for substrate binding and activity.
- The presence of Mn2+ inhibits 3'-exonuclease activity but stimulates endonuclease activity.
- Rrp44's N terminal PIN domain is vital for exosome core structure linking.
- Rrp44-endo mutation maintains the endonuclease activity of Rrp44.
- The exosome's exonuclease activity is critical in RNA processing.
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Description
Explore the intricate roles of Rrp44 in the yeast core exosome. This quiz delves into the exonuclease and endonuclease activities of Rrp44, their developmental impacts, and the synthetic lethality involved in its functions. Test your understanding of RNA processing and pre-rRNA maturation in yeast.
