Closing Thoughts on My Summer Research Project

Hello There!

With my summer projects complete, and with an adequate time for reflection about my results and the future directions of my projects afforded, I now feel ready to complete my last post of the season.  From the data gathered from the autophagosome, lysosome, autolysosome, and L3CB experiments, I have seen first hand some of the fascinating cellular effects that MK-STYX exerts.  With this protein established as an important autophagy player in our in vitro studies, exciting new experimental directions beckon in other organismal models.  In the future, I will explore how autophagy transcription factors, the activators of specific genes in DNA, are influenced by MK-STYX.  Following this, I will investigate the known role MK-STYX plays in clearing misfolded proteins, or protein aggregates, from cells.  I believe this represents an important experimental direction as these variousprotein aggregate types are major factors in devastating diseases like Huntington’s Disease, ALS, and Alzheimer’s.

[Read more…]

Further Development of My Project

Hello Readers!

The investigation into MK-STYX’s link with the autophagosomal marker protein LC3-II has yielded fascinating results, and adds a new dimension to our hypothesis that MK-STYX functions as a significant regulator of autophagy.  While we see how MK-STYX could promote the degradation of components already identified by the cell for clearance, I believe it is worth investigating how MK-STYX could impact upstream of this.  Specifically, I will look into if MK-STYX may affect the specific proteins and organelles that are “tagged” for digestion intracellularly and how exactly MK-STYX achieves this target marking.  Our lab carried out previous work wherein we identified MK-STYX as a clearer of stress granules and protein buildups known as aggressomes.  Because autophagic processes are a primary means of effecting the clearance of these cellular features, I wish to explore how MK-STYX could recruit the autophagy effectors like autophagosomes and lysosomes to these aggregations.

[Read more…]

The Next Phase of My MK-STYX/Autophagy Summer Research

Hello Everyone!

After concluding some exciting work on MK-STYX and its impacts on autophagy organelles, I will now turn my attention to how our lab’s protein of primary interest, MK-STYX, may impact the production of the essential autophagy component LC3-II.  This protein is essential to the generation of the signature autophagy vesicle known as the autophagsome, so quantifying a change in its cytosolic expression after activating MK-STYX would strengthen our hypothesis that posits MK-STYX as a significant player in the autophagy pathway.  Additionally, I will be working closely with my inspirational autophagy project teammates Emily Pickering and Y-Nhui Bhui to characterize the nature and extent of MK-STYX’s activation of autophagy-promoting transcription factors.  We will analyze how the localization of these crucial polypeptides may shift using Cell Profiler computer algorithms manual scoring.  I will post an update in a few days about my progress with the aforementioned topics and my research plans for the rest of the summer.

[Read more…]

Research Progress Update

Hello All!

We are now a few weeks in to the Summer 2018 research session and there are many developments I would like to relate.  After consultation with my lab mentor Dr. Shanta Hinton and lab alumnus Patrick Christian, I have added a computer-driven element to my project.  Using the Broad Institute’s open-source program CellProfiler, I have been able to characterize the effects of our protein of interest, MK-STYX, in thousands of different cells across more than a dozen different parameters.  This has helped further visualize MK-STYX’s impacts on autophagy factors like autophagosomes (cellular garbage trucks) and autolysosomes (cellular recycling centers).  Additionally, I have been utilizing the genetically refined BacMam virus to gain important insights into the inner workings of the cell.  This autophagy sensor lights up various points of interest and allows for them to be compared, counted, and analyzed across different experimental groups.  It has been an excellent time in lab so far, and with the help of Dr. Hinton and my brilliant lab mates, I expect to see many more intriguing research findings this summer.

Exploring the Roles of Pseudophosphatase MK-STYX in Autophagy

Exploring the Roles of Pseudophosphatase MK-STYX in Autophagy

 

Cells carry out an incredible variety of processes to keep themselves and the greater organism they comprise functioning efficiently and healthily. As with most complex systems, cells possess a means of eliminating unwanted components and reusing or recycling the individual parts of these superfluous components into important constituent parts like amino acids and nucleic acids. This cellular recycling system is known as autophagy, and it permits organisms and their various cell types to remove redundant biomolecules [1]. In addition, autophagy allows cells to sustain themselves in times of starvation or stress by breaking down complex organelles and proteins into energy and basic biological building blocks. Unsurprisingly, autophagy is highly relevant to both healthy and diseased states, because a baseline level of autophagy is essential to an organism’s physical wellbeing. Conversely, a grossly dysregulated autophagic process affects and mediates pathogenesis and progression of cancers, neurodegenerative diseases, and metabolic disorders [2]. The pseudophosphatase MK-STYX and its roles in cellular processes represent the primary research focus of the Hinton Lab.  As a pseudophosphatase, MK-STYX lacks the catalytic ability to remove a phosphate group from proteins, but can bind targets with its pseudophosphatase domain and protein-interacting CH2 domain.  The importance of pseudophospahtases has only recently been accepted, thus MK-STYX and the other proteins in this group populate an exciting forefront of scientific inquiry.  Previous findings from our research lab have demonstrated that MK-STYX activity helps clear stress granules (made of translationally halted mRNA and associated proteins) from cells [3].  The two pathways that clear stress granules are the autophagy and ubiquitin protease pathways.  Based on recent studies conducted by our lab that showed MK-STYX altering the activity and expression levels of autophagy proteins, I decided to focus my research efforts on further characterizing MK-STYX’s roles in this critical intracellular homeostatic process.

[Read more…]