Autophagosomal Pool Flow

Since AMPK sites in ULK1 are conserved, the group also examined if the two proteins work together in autophagy conservation in C. elegans using reporter assay and histogram plots.

The PI3Ks are linked to variety of different signaling pathways, but each version of the lipid enzyme has a similar PIK core region [2]. This region of the PI3K is composed of three essential parts: a C2 region, a helical region, and a catalytic region; it should be noted that in some forms of the PI3Ks these regions are split up on different subunits (refer to Figure 1 below) [2]. From these three structural components, the PI3K is able to carry out its important activating role and create the three products listed above. These products are noted to remain near the plasma membrane- on the cytosolic side- and to have their own biding sites [3]. The binding sites on the PI3K products are indications of the signaling pathway, and therefore attract other signaling proteins, like the serine-threonine kinases PKB and PDK1 [3]. It is also important to mention that the PI3K signal and its products can both be terminated by removing the phosphate added to the inositol ring of the phosphoinositide; this is done by the lipid phosphatase known as PTEN [3]. A termination step is important in the regulating activity of PI3Ks, and therefore in regulating the activities of the cell. From the medical field perspective, PI3Ks are important since they are linked to crucial pathways for insulin regulation and cell growth.

MicroRNA’s can affect the gene expression by preventing their translation onto protein. This blocking function lead to enabling the cells to adapt to change condition.

microRNAs (miRNAs) are small regulatory RNAs that can regulate the gene expression of a large number of target messenger RNAs (mRNA){Bartel:2009fh}. miRNA biogenesis in most cases begins with RNA polymerase II transcription of a long primary miRNA (pri-miRNA) containing one or more hairpin structures that are processed by the nuclear RNase III endonuclease Drosha and the double stranded RNA-binding protein, DiGeorge Syndrome Critical Region Gene 8 (DGCR8){Lee:2003hq} (Figure 1.4). This cleavage generates a 70-nucleotide stem loop known as the precursor miRNA (pre-miRNA), which is actively transported into the cytoplasm by a complex of exportin-5 (XPO5) and Ran-GTP{Yi:2003gz}. The pre-miRNA is then cleaved by a second cytoplasmic RNase III endonuclease, DICER, which acts in complex with the RNA-binding protein TAR (HIV-1) RNA Binding Protein 2 (TARBP2) to generate an

In the fluorescence experiment, the anti-GFAP (Glial fibrillary acidic protein) antibody binds to the GFAP protein in the tissue. Putting on the secondary antibody helps visualise where the GFAP is and acts as an amplifier. The second antibody has an added fluorescent probe and will glow when an electromagnetic energy of a certain wavelength shines on it. This will help see where the GFAP is bound and the type of cell it is in. In this case, in the brain tissue, will be

It has long been known that AF induces electrical and structural remodelling, and the phenomenon that AF begets AF is accepted as an explanation for the progressive nature of lone AF from a paroxysmal to sustained condition, but little has been known about the underlying mechanisms. However, now that knowledge is beginning to emerge as to how these changes can occur, the issue presenting itself is how to utilise this. In order to do this, the limitations of animal models, difficulties of miRNA profiling, and the implications of attempting to alter the balance of miRNA expression must be considered prior to further consideration as therapeutic targets.

exRNA was submitted to the Beijing Genome Institute (BGI, Shenzhen, China) for cDNA synthesis and RNA-Seq. Approximately 100 ng cDNA from each strain was each subject to illumina deep sequencing using BGISEQ-500 technology. A total of 27,084,174 clean sRNA reads was obtained from the cin1 mutant and 27,402,307 clean reads were obtained from WT (Table 1). The length of sRNA is between 18 nt and 30 nt. The initial sRNA annotation provided by BGI suggested that approximately 96% sRNA sequences were mapped to the available sRNA database including miRBase and Rfam. Further annotation, prediction, expression, screening of differentially expressed sRNA, and miRNA target prediction will be performed under the direction of Dr. Taylor. Using this methodology, we will determine the role of Cin1 in secretion and uptake of sRNA.

Background: High blood glucose fluctuation (BGF) contributes in chronic complications by promoting the dysfunction of vascular endothelium in diabetes. MicroRNA (miR)-34 family is known for its roles in the pathophysiology of diverse diseases including type 2 diabetes. However, the action of BGF on the expression of these miRs in diabetes is unknown. Herein, we aimed to investigate the effects of BGF on the pathogenesis of diabetes and the possible involvement of miR-34 family in vivo.

To potentially address the functional significance of miRNA, we have further evaluated the miRNA-mRNA targets by using

Type III and IV are vesicular based pathways 10. Type III includes “autophagy based secretion”, while type IV is for “proteins that

The oxidative stress can cause cell damage, resulting in inflammation, which has also been found to be a major contributor to disease progression of GM2 gangliosidosis [17]. In this study, we identified elevation in glutathione pathways, which plays a pivotal role in responses to oxidative stress. Another evidence of inflammation is reduced levels of arachidonic acid, an omega-6 fatty acid, in brain samples of SD mice. Oxidation of arachidonic acid can generate leukotrienes, a family of eicosanoid inflammatory mediators produced in leukocytes, and thus promote inflammation. The increased energy requirements can also activate autophagy and protein catabolism, which have been found in MPS I and MPS VII mice [13]. In this study, we found increased levels of amino acids, amino acid derivatives and dipeptides, indicating increased protein catabolism. Increased requirements of energy and raw materials can also activate lipid metabolism and carbohydrate metabolism, manifested by decreased adiposity, a common observation in many lysosomal diseases [18-20]. In addition, the enlarged lysosome and distended cells due to abnormal accumulation requires increased membrane synthesis, which can also affect lipid metabolism. Our previous proteomic analysis [21] also identified abnormality in the cytoskeleton system, which can be partially attributed to altered cellular architecture due to storage accumulation. Collectively, we show here that the energy imbalance caused by the lack of

miRNA’s have been found in many organisms even including plants. They have been shown to regulate various processes , some as fundamental as development, and differentiation. Their role has been found to be a negative regulator of gene expression. miRNA’s bind to the transcribed mRNA and cause it to be degraded, thus inhibiting the production of the gene product. It is not surprising that in cancer, miRNAs could either inappropriately repress anti-tumor mechanisms (such as steps in the apoptosis pathways) or loss of the miRNA

The mechanisms of apoptosis are highly complex involving an energy dependent cascade of molecular events. Research

The miRNAs control gene expression post-transcription through different mechanisms such as repression at the initiation step by repression by preventing 60S subunit joining, deadenylating, proteolysis, and slow elongation or ribosome drop-off. This mechanism regulates mRNA translation or stability in the cytoplasm via non-perfect pairing with target mRNAs. Usually involving a seed pairing of just six to eight nucleotides in length cause degradation of target RNAs by the RISC complex in the case of perfect complementarity with the target site—the phenomenon known as RNAi. It is estimated that approximately one-third of human protein-coding genes are controlled by miRNA.

My project work is focused on understanding the role of non-coding RNA – miRNAs regulating cellular proliferation during oncogenic transformations. miRNA targeting mRNA regulates gene expression post transcriptionally and plays an important role in tumor progression. To identify differentially expressed miRNAs in Osteosarcoma cell lines which have different proliferation rates, I have screened miRNAs from relevant microRNA datasets and upregulated and downregulated microRNAs were shortlisted and their expression were analyzed in the respective OS cell line by real-time RT PCR. Potential targets for miRNAs were screened for cell cycle and DNA replication genes by target scan and KEGG database. Downregulated miRNAs were overexpressed in OS cell lines by transfecting the synthetic mimics/ inhibitors and checked the expression levels of the target protein at RNA level by qRT PCR and at protein level by Immunoblotting. Further cell