VIPSolutionsV4 The image below shows the results of a gel shift assay. Each column (i.-vi.) contains a sample with the same radioactive oligonucleotide probe complementary to a region of DNA which is suspected to be an enhancer sequence. This DNA is known to bind several transcription factors of the same family (HFN). Samples ii. - vi contain whole nuclear extract (N.E.) isolated from hepatocytes. A gel shift is performed to determine which member/s of the HNF family of transcription factors bind to this DNA sequence in question. To this end, some of the mixtures contain antibodies specific for distinct HNF members (written along top). § 57322 ee”. 111% Based on this data, which of the following transcription factors listed are bound to the region of DNA in question? NF-4a and HNF-1a HNF-1a and HNF-1B HNF-4y and HNF-1B.. HNF-1a and HNF-4y all HNF factors bind to this DNA NE. TTT Sh SR SE 4 LA - w v “.
Question:
V4 The image below shows the results of a gel shift assay. Each column (i.-vi.) contains a
sample with the same radioactive oligonucleotide probe complementary to a region of
DNA which is suspected to be an enhancer sequence. This DNA is known to bind several
transcription factors of the same family (HFN).
Samples ii. - vi contain whole nuclear extract (N.E.) isolated from hepatocytes. A gel
shift is performed to determine which member/s of the HNF family of transcription factors
bind to this DNA sequence in question. To this end, some of the mixtures contain
antibodies specific for distinct HNF members (written along top).
§ 57322
ee”. 111%
Based on this data, which of the following transcription
factors listed are bound to the region of DNA in question?
NF-4a and HNF-1a
HNF-1a and HNF-1B
HNF-4y and HNF-1B..
HNF-1a and HNF-4y
all HNF factors bind to this DNA
NE. TTT Sh SR SE 4
LA - w v “.
This is AI generated Response.
Asked by: Nikita Gaikwad
Created at: 2025-06-19 00:53:24
Asked by: Nikita Gaikwad
Created at: 2025-06-19 00:53:24
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Let's break down the gel shift assay results:
- **Lane i (-)**: Contains DNA + nuclear extract (N.E.), no antibody. Shows a shifted band, indicating that a protein from the extract binds the DNA.
- **Lane ii (HNF-4α antibody)**: Shifted band moves to a higher position (supershift), indicating HNF-4α binds the DNA.
- **Lane iii (HNF-4γ antibody)**: No supershift, same as lane i, suggesting HNF-4γ does **not** bind the DNA.
- **Lane iv (HNF-1α antibody)**: Supershift, indicating HNF-1α binds the DNA.
- **Lane v (HNF-1β antibody)**: No supershift, suggesting HNF-1β does **not** bind the DNA.
**Interpretation:**
- Supershift occurs **only** with HNF-4α and HNF-1α antibodies, which means these proteins are binding to the DNA.
**Correct answer:**
> **1.) HNF-4α and HNF-1α**
Question:
explain more
This is AI generated Response.
Asked by: Nikita Gaikwad
Created at: 2025-06-19 00:53:45
Asked by: Nikita Gaikwad
Created at: 2025-06-19 00:53:45
Final Answer:
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Certainly! Let's analyze the gel shift (EMSA) data step-by-step to understand which transcription factors bind to the DNA sequence in question.
---
### Overview of the Gel Shift Assay Data:
| Lane | Description | Observation | Interpretation |
|--------|----------------|----------------|----------------|
| i. | DNA + nuclear extract (no antibody) | **Shifted band present** | Proteins in the extract bind the DNA, causing a shift. |
| ii. | DNA + nuclear extract + HNF-4α antibody | **Supershift (further shifted)** | HNF-4α binds the DNA; antibody binding causes a supershift. |
| iii. | DNA + nuclear extract + HNF-4γ antibody | **No supershift** | HNF-4γ does not bind the DNA. |
| iv. | DNA + nuclear extract + HNF-1α antibody | **Supershift** | HNF-1α binds the DNA. |
| v. | DNA + nuclear extract + HNF-1β antibody | **No supershift** | HNF-1β does not bind the DNA. |
---
### What is a supershift?
- When an antibody binds to a DNA-protein complex, it increases the size of the complex, resulting in a **supershift**—a band that migrates slower (higher position) on the gel.
- **Presence of a supershift indicates that the specific protein recognized by the antibody is part of the DNA-protein complex.**
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### Interpretation:
- **HNF-4α**: The supershift in lane ii indicates HNF-4α binds the DNA.
- **HNF-4γ**: No supershift in lane iii suggests HNF-4γ does not bind.
- **HNF-1α**: The supershift in lane iv indicates HNF-1α binds.
- **HNF-1β**: No supershift in lane v suggests HNF-1β does not bind.
### Conclusion:
- **HNF-4α** and **HNF-1α** are the transcription factors binding to the DNA in this assay.
- The other HNF family members tested (HNF-4γ and HNF-1β) do not bind under these conditions.
---
### Final answer:
**HNF-4α and HNF-1α** are the transcription factors bound to this DNA region.
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