Cell and Molecular Biology

Cell and Molecular Biology concluding Name Hadeel Binomar30 pts. Protein misfolding can be an aspect of several different kind-hearted disorders, including cystic fibrosis, Alzheimers disease, and atherosclerosis. M whatever times, the misfolded protein is a membrane protein.In fact, a pillow slip of diabetes insipidus results from a mutation in the G-protein-coupled vasopressin 2 sense organ that prevents the protein from reservation it to the cadre membrane A) Describe how this type of receptor would normally beat targeted to the membrane (from the beginning of translation) and B) indeed propose one mechanism by which the mutation could cause a lack of proper targeting.One of the close to important proteins target is G protein-coupled receptors, several signaling mechanisms depend on this type of receptor to change both internal and external stimuli to the intra boothular responses. Basically, one of the G couple receptor subfamilies is G-coupled Vasopressin-2- Receptor (V2 R), and this receptor is field of study through a strict quality control process at the endoplasmic reticulum, which presents the only correctly folded protein to gets through the secretory pathway.The elementary process of the V2 receptor is to activate the attached G protein that bound to the ? subunit then phosphorylated to GTP. The G protein couple receptor then activates the enzyme adenylate cyclase that catalyzes the reaction in the ER and forms bivouac from ATP. After that, cAMP acts as a second messenger and activates a protein kinase that phosphorylates the constitutional membrane proteins on the stall develop.Moreover, the secretory pathway organelles and the plasm membrane both atomic number 18 first introduced into the Endoplasmic reticulum, and the co-translationally proteins that can cross the ER be synthesized by the ribo slightly first then beats by chaperones to gets moves to the ER surface victimisation GTP that allows them to move toward the receptor t hen release it.The soluble proteins and the integral membrane proteins as I mentioned above, can be targeted through the ER and then translocated by the same mechanism.Further, several mutations occurring in the transmembrane region which impact the structure of the protein. These be multiple mutation sites, such(prenominal) as mutations occurring in the aminic acid residues which was acting as a causative agent for humanity disease.Also, there be other mutations occurring on the single site position that will affect the translation mechanism and cause more human diseases like cystic fibrosis, Alzheimers disease, and atherosclerosis that proves the function of V2R protein which plays important occasion in the translation during the protein folding process.One of the mechanism that may a reason for lacking the proper target protein is when the mutations of the CFTR constituent occurring and affect the function of the chloride ion channels and cause defect in the protein seq uences which lead to the production of diseases and misfolded of the proteins that are unable to recognize their functional destinations.Otherwise, Lack of the determine signals is another issue that prevents the protein from getting into the surface of the ER, also called the non-stop decay electric cellular pathway, because lack of this stop signals prevents mRNA from synthesis and translate the proteins, these distribute as a plosive mutation that inhibits the essential stop codons. 30 pts. Describe the try shown in figure 3 from the paper we discussed in class (Miller et al, 2003). (A)In your description, consider the following questions Why did they do it? How did they do it? What did they learn? (B) Diagram the results that you would seem to see in Lanes T, 1, 2, 3, and 4, if the amino acid signal DID in the protein Gap1p was mutated to stochastic amino acids and tell why and (C) Give two possible (different) results that talent occur if the amino acid signal LxxLE in B et1p was mutated to the amino acids DID (which are the signal in Gap1p).Diagram the results expected in Lanes T, 1, 2, 3, and 4 and develop why you predicted this result for each case.left20840701.A001.A4467225201739500The experimentation was performed to study the role of freight rate binding do of import of Sec24p in the process of protein sorting. To perform this study, both edition and wild types subunit Sec23/24p and Sec23/24L616W were harvested from microsomal membranes.The immunoblotting assay performed to quantify cargo molecules using radio-labeled secondary antibodies. Comparison of the mutant subunit with wild type illustrated disrespect of some molecules in mutant one, these molecules were cargo protein molecules. While some of them are packed in a way that similar to the wild-type. It meant that there were some signals that remained unaffected although mutation was there.Unexpectedly, it was found that in these unaffected molecules of mutation in Sec23/24p, packag e was better than the wild type. Further, they found the proteins that were highly affected had Bet1p and Gap1p/Sys1p chimera, because they tout ensemble depending on the Sys1p di-acidic of COPII vesicles. If the amino acid signal DID in the protein Gap1p was mutated to random amino acids, the resulting bands would be seen as in figure 1.A because the amino acids mogul be present in all the lanes. And Gap1p is required for di-acidic motifs to fuse together with the COPII vesicles at the mutated domain. Also, Sys1 peptide is not involved in Sec 23/24p because its mutation is not contained in a di-acidic motif. If the signal LxxLE was mutated to the DID, the expected Bet1p packaging would be shown in figure 2.A as not be seen in lanes containing Bet1p because the amino acid signal LxxLE does not exit later mutating to amino acids DID. The second probability for the Bet1p would be expected to bind with the vesicle for packaging when it interacts with di-acidic motifs which is need ed as shown in figure 2.B290512515621000581025113030004781550412752.B002.B6286501346202.A002.A20 pts. Explain the experiment shown in Figure 5C from the Shen et al. (2018) paper on the phosphorylation of CDC25 that we discussed in class.In checkition, be sure to also address the following A) why did they do this experiment (the central issue) B) what did they learn and C) what is another control experiment they could have do? D) Would these results change if you added a constitutively active form of LKB1 to the reaction? If so, how and why? A) Overall, the main purpose of this paper is to explain how the division of cells is highly regulated such that cells that fail to pass some specific stage-based tests cannot advance to the proceeding stages.In this particular experiment, HeLa cells were subjected to chemicals that are known to activate the enzyme AMPK. Also, the main catabolic processes that are involved to generate nada for cells to regeneration from G1/G2 were determined by the application program of radiochemical approaches, the experiment required to approve how highly conserved cellular energy sensor can significantly delay mitosis entry and energizing of AMP-activated protein kinase (AMPK).Further, defecate 1 family inactivates the cell cycle G2/M Phase which is controlled by Cdc3/cyclical B (mitotic cyclin-dependent kinase complex). They found that AMPK-dependents phosphorylation of CDC25C arranges a metabolic control point for the M- bod transition and the cell cycle var. G2.B) Also, they learned that suppression of Wee 1 or acute induction partially reinstates mitosis ingress in the context of activated protein kinase (AMPK). This experiment showed that when Phosphorylates Cdc25 is in a distributive and disordered state, it results into ultra-sensitivity in protein phosphorylation. C) For another control experiment, they might try exposing the U2OS (cell line) clones conditionally exhibiting Cdc25A etoposide.That will protagonist to test whether Cdc25A degradation is significant for the G2 control point or not.D) If they add an active form of LKB1 to the reaction, the results will not change, because in cells LKBI activity inhibits AMPK activation in response to different stimulations. Also, LKBI is lost upon consistent isolation and because no effects of phosphates observed.20 pts.Briefly describe the role of cyclin-CDK in the cell cycle and give an example of positive and negative control in this system. Also, describe how cell cycle regulation links to the stimulation of caspase-mediated cell death at the molecular level. In the beginning, the cell cycle depends on many an(prenominal) basic factors that control the regulation process starting from the signal organization molecules, growth factors and the Cyclin Dependent Kinases enzymes include the checkpoints which control the transition process surrounded by the cell cycle phases by binding to the cyclin proteins CDKs then phosphorylate other proteins to move out from one phase to another.The role of transcription factors is to turn on the signals for gene expression, DNA replication, and cell divisions. As an example of CDKs, cyclin-dependent kinase 1 CDK1 is a cell division cycle protein homolog 2, that has a primary role in human genus Cancer cells because CDK1 rather than any types of CDKs is fatal to the mutated version of MYC- dependent cancer that leads to a depletion of oncogenes like (Fos and Jun) in human cancer cells.Fos and Jun are feature forms of the transcription factor called AP1 that activates the delayed response genes such as cyclin D and CDK4. Recent studies reveal that the reason for the MYC breast cancer cells duplation is targeting CDF1 exhibit any other CDKs cell lines. Also, CDK1 inhibition can control and target the cancer cells in human and both phosphorylation and expression of MYC during the cell cycle process.To novitiate intracellular signaling pathways and draw the cell cycle entry, mitogens sub stance bind to the cell surface receptor with the activation form of GTPase Ras that activates present kinase cascade. That will lead to the expression of encoding gene of the transcription regulative protein like MYC. Moreover, E2F transcription factor is a target for cyclin D and CDKs that stimulate proteins expression to initiate S phase, also E2F regulated by the tumour suppressor gene gene Rb (Retinoblastoma protein).At early G1 Phase, Rb protein unite with E2f to form the histone deacetylases protein that remains chromatic condensed, then cyclin kinase phosphorylates Rb protein, causing it to separate from E2F. That allows recruitment of histone acetylates, which decondense the chromatin and helps transcription complexes to form in G1 and S phase and to form a positive feedback.For the negative control, if there are no growth factors present to stimulate the synthesis of Cyclin D in the peeled cell, Rb phosphorylated will turn off, and Rb will rebind to E2F, then the cel l cycle will stop and that will lead to many negative results like prevent it to bind with DNA, or DNA disability and cells will return to the G0 phase.Apoptosis is a consequence of DNA disparage if the damage is not repaired, the cell cycle will divert towards apoptosis, otherwise, if the cell has DNA defects and fail to undergo apoptosis, that will change to the cancer cell. G1 phase regulators such as P53 and E2F are essential to promote the cell regulations and top any damage or abnormal changes during the cell cycle.P53 has a primary function to prevent any mutation in DNA during cell progression, so any defect in P53 will lead to cancer, and its commonly mutated in cancer cells. Further, Rb protein considers as a tumor suppressor and also promotes apoptosis. Additionally, most human cancers have inactive Rb protein, every mutated Rb or the non-phosphorylated (inactivated form) of Rb protein