RNA Interference Video Lessons

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Problem: Members of the Argonaute protein family play key roles in RNA interference pathways (RNAi). RNAi lowers gene expression in response to double-stranded RNA (dsRNA) whether endogenous (e.g. pre-miRNA) or exogenous (e.g. viral genomes). The RNA process is initiated when Dicer, an RNase IlI family enzyme, cleaves dsRNA into short duplexes (~21 base pairs), each strand possessing a single-stranded, two-nucleotide overhang on its 3'–end and a phosphate on its 5'–terminus. These dsRNAs (siRNAs or miRNAs) are then loaded onto Argonaute (Ago).Once engaged by Ago, one of the siRNA or miRNA strands, the passenger strand, is cut and discarded. The remaining single-stranded RNA (ssRNA) is termed the guide strand. Ago acts as the scaffold for the recognition of complementary target sequences by the guide strand. Additionally, as part of the RNA-induced silencing complex (RISC), Ago can catalyze the cleavage (slicing) of target mRNAs.Identify the domains of argonaute labeled 1-3a. PAZ domain (1), Middle domain (2), N-terminal domain (3) b. PIWI domain (1), Middle domain (2), PAZ domain (3)c. Middle domain (1), PIWI domain (2), N-terminal domain (3)d. PAZ domain (1), PIWI domain (2), N-terminal domain (3)Which Argonatue domain contains the active sitea. PAZ domainb. N-terminal domainc. Middle domaind. PIWI domain

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Argonaute Protein Structure:

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Problem Details

Members of the Argonaute protein family play key roles in RNA interference pathways (RNAi). RNAi lowers gene expression in response to double-stranded RNA (dsRNA) whether endogenous (e.g. pre-miRNA) or exogenous (e.g. viral genomes). The RNA process is initiated when Dicer, an RNase IlI family enzyme, cleaves dsRNA into short duplexes (~21 base pairs), each strand possessing a single-stranded, two-nucleotide overhang on its 3'–end and a phosphate on its 5'–terminus. These dsRNAs (siRNAs or miRNAs) are then loaded onto Argonaute (Ago).

Once engaged by Ago, one of the siRNA or miRNA strands, the passenger strand, is cut and discarded. The remaining single-stranded RNA (ssRNA) is termed the guide strand. Ago acts as the scaffold for the recognition of complementary target sequences by the guide strand. Additionally, as part of the RNA-induced silencing complex (RISC), Ago can catalyze the cleavage (slicing) of target mRNAs.

Identify the domains of argonaute labeled 1-3

a. PAZ domain (1), Middle domain (2), N-terminal domain (3) 

b. PIWI domain (1), Middle domain (2), PAZ domain (3)

c. Middle domain (1), PIWI domain (2), N-terminal domain (3)

d. PAZ domain (1), PIWI domain (2), N-terminal domain (3)


Which Argonatue domain contains the active site

a. PAZ domain

b. N-terminal domain

c. Middle domain

d. PIWI domain

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