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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 gene is embedded within a CG island.
The researcher uses a specialised laboratory technique to examine DNA methylation in the promoter region. What do you hypothesise the researcher will find? (1 mark)
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 promoter region in cardiomyocytes vs fibroblasts (i.e. skin cells).
Use the information provided above to explain why the gene is actively expressed in cardiomyocytes but not fibroblasts. (2-3 sentences) (2 marks)
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?
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)
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.
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
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)
What structural motifs can you identify in the interactive ribbon model of the acyl-CoA dehydrogenase homotetramer? (select all that apply)
Which of the following techniques was likely used to elucidate the three-dimensional structure of the acyl-CoA dehydrogenase homotetramer?
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.
Examine the western blotting results shown in the image above (A, B, C). Which result would you expect to obtain for this experiment?