What key pair of cell cycle experiments were done using RNase? What are TWO key findings from this experiment? (hint: for molecular cell biologists? For evolutionary biologists?)
What key pair of cell cycle experiments were done using RNase? What are TWO key findings from this experiment? (hint: for molecular cell biologists? For evolutionary biologists?)
Homework Answers
Answer:-
1) FLOW CYTOMETRY FOR DNA ANALYSIS
- The DNA needs to be stained and permeabilized.
- This can be achieved by treated the cells with alcohol (ethanol) or by any detergent. (Sodium Laurel Sulfate)
- Permeabilization enhances interaction of DNA with nuclear content and RNAses are added to ensure that RNA does not get the stain after the treatment.
- So, utilization of RNAase A is to enhance the specificity of staining with respect to DNA during flow cytometry studies carried out to analyse DNA.
- Propidium iodide is the intercalating agent used to stain the DNA.
- Cells have the G1 phase that determines the regulatory control over cell cycle and is associated with pathogenesis or cancer formation.
- Flow cytometry is specifically used to analyse such regulatory changes. This includes quantification of the DNA replication process and to analyse the Phase where replication initiation decision is being undertaken. Role of RNAses is to just enhance specificity for DNA determination by eliminating RNA source.
2) RNAse H determination in retroviruses as key for evolution
- RNAses H are found as molecules associated with reverse transcriptase that are involved in viral replication via RNA-DNA hybrid to transfer the required sequence into the host.
- It plays an important role in cell cycle regulation in terms of replication events and is considered to have evolved from the ribozymes.
- RNAse H is also associated with activity of the transposable elements.So, basically this mechanism is associated with changes in the sequence of genome of HIV virus due to which specific drugs targeted towards the genes for virulence becomes ineffective. So, basically a research was carried out to inhibit RNAse H activity that is assocaited with activity of transposable elements.
- Apart from this, it mediates regulation of viral replication and thus maintains the balance between transposable elements and replication by means of reverse transcription.
- So, the role was to study the RNAse H interaction with respect to the cell cycle of virus being a part of reverse transcriptase complex in order to develop a drug specifically targeting RNAses for altering its regulatory control over cell cycle.
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What key pair of cell cycle experiments were done using RNase? What are TWO key findings from this experiment? (hint: for molecular cell biologists? For evolutionary biologists?)
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