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MCB2011 - Molecular biology and the cell - S1 2025

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The ACTB

gene encodes an actin protein that is ubiquitously expressed in all human

cells. As shown in the schematic below, the promoter region of the

ACTB

gene is embedded within a CG island.

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The researcher

uses a specialised laboratory technique to examine DNA methylation in the

ACTB

promoter region. What do you hypothesise the researcher will find? (1 mark)

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The ACTC1

gene encodes an actin protein that is only expressed in cardiomyocytes (i.e.

cardiac muscle cells). The schematic below shows the histone modifications that

are present in the

ACTC1

promoter region in cardiomyocytes vs

fibroblasts (i.e. skin cells).

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Use the

information provided above to explain why the

ACTC1

gene is actively expressed in

cardiomyocytes but not fibroblasts. (2-3 sentences) (2 marks)

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The biomedical

researcher discovers that the defective acyl-CoA dehydrogenase subunits in the HEP-M2

cell line are targeted for proteasomal degradation. Which post-translational

modification is likely responsible for targeting acyl-CoA dehydrogenase to the

proteasome?

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Explain why the mutation in the HEP-M2 cell line completely disrupts the

assembly of the acyl-CoA dehydrogenase homotetramer, whereas the mutation in

the HEP-M3 cell line has no impact on the assembly or structure of the

homotetramer. (2-3 sentences) (2 marks)

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The import of acyl-CoA dehydrogenase into the mitochondrion is defective

in cell line HEP-M1. The researcher used subcellular fractionation and western

blotting to detect acyl-CoA dehydrogenase in different subcellular

compartments. The western blotting results for normal healthy hepatocytes and the

HEP-M1 cell line are summarised in the figure below.

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Which of the

following mutations (A, B, C) is most likely present in the HEP-M1 cell line?

Use the western blotting results and your knowledge of the mitochondrial

protein import process to justify your response. (2-3 sentences) (2 marks)

A. The acyl-CoA

dehydrogenase mitochondrial signal sequence is mutated

B. The

receptor protein in the TOM complex is mutated

C. The

translocation channel in the TIM23 complex is mutated

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Acyl-CoA dehydrogenase is encoded in the nuclear genome, synthesised by

free ribosomes in the cytoplasm, and then imported into the mitochondrial

matrix. Acyl-CoA dehydrogenase is subject to which type(s) of intracellular

transport? (select all that apply)

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What structural motifs can you identify in the interactive ribbon model

of the acyl-CoA dehydrogenase homotetramer? (select all that apply)

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Which of the following techniques was likely used to elucidate the three-dimensional

structure of the acyl-CoA dehydrogenase homotetramer?

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A researcher extracts protein from cultured hepatocytes (i.e. liver

cells) and treats the protein sample with sodium dodecyl sulphate (SDS) and

β-mercaptoethanol (BME). The protein sample is loaded onto a polyacrylamide gel

and a western blot is performed as per the steps described in the relevant online

learning module (OLM 6.2). The researcher uses a primary antibody that specifically

binds to the N-terminal region of acyl-CoA dehydrogenase.

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Examine the

western blotting results shown in the image above (A, B, C). Which result would

you expect to obtain for this experiment?

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