Research Group of Dr Marilyn Resh, Sloan Kettering Institute

Research Group of Dr Marilyn Resh, Sloan Kettering Institute

Marilyn Resh is a member of the Cell Biology Program at the Sloan Kettering Institute, Memorial Sloan-Kettering Cancer Center. The goal of her laboratory's research is to understand how fatty acylation influences the structure and function of membrane-bound signaling proteins, which regulate a number of cellular activities, such as intracellular protein trafficking, intracellular signal transduction, and even virion formation. The group’s focus is on members of the Src family, a group of myristoylated membrane-bound tyrosine protein kinases.

Studies by the Resh lab established that membrane association of Src results from synergism provided by hydrophobic insertion of myristate into the lipid bilayer and electrostatic interaction of the positively charged amino acids with negatively charged head groups of acidic membrane phospholipids. This "myristate + basic" motif is also found in the Gag proteins of many retroviruses. Their laboratory showed that the myristate + basic domain mediates plasma membrane targeting of HIV-1 Gag, thereby allowing Gag to function in the formation and budding of virions. They are currently studying the molecular mechanisms involved in retroviral particle formation at the plasma membrane.

Marilyn did her Ph.D in the Department of Biochemistry and Molecular Biology at Harvard University and performed her post doctoral work in the Department of Cellular and Developmental Biology also at Harvard. Marilyn then became Assistant Professor in the Department of Molecular Biology at Princeton University. In 1991, she moved to the Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, where she is a Member and Professor in the Cell Biology Program. In addition, Marilyn currently has a joint appointment as Professor of Cell Biology and Genetics and Professor of Biochemistry at Cornell University Graduate School of Medical Sciences/Weill Graduate School.

The Resh lab uses Volocity in combination with timelapse confocal imaging to understand the trafficking of c-Src within cells. They use Volocity to view large 3D timelapse datasets and to track the intracellular movement of vesicles containing c-Src-GFP that originated from the plasma membrane. Now that they have established a system for tracking c-Src vesicles using Volocity, they will be able to further test the properties of c-Src movement in the presence or absence of other proteins or drugs which perturb cellular trafficking pathways.

The Resh lab will be among the first to take advantage of the important new Volocity Site License recently installed at the Sloan Kettering Institute. This provides access to Volocity, administered via the Imaging License Server software. This impressive installation provides network access to Volocity for all the researchers at Sloan Kettering. The program can be installed on any number of suitable Macs and PCs, and users can log into the system remotely to access all the high performance 3D and 4D imaging features that Volocity provides.

Marilyn says "Volocity has provided my laboratory with sensitive, high resolution tools to quantitate intracellular protein trafficking in three dimensions. These measurements will enable us to determine the role of key regulatory proteins on the movement and signaling capacity of c-Src and other important signal transducers. The new Imaging License Server means that my lab will have immediate access to the software tools that we require for our work with quantitative imaging analysis."

More detailed information about the lab’s research projects is explained on the laboratory web site.

COS-7 cells expressing cSrc protein fused to monomeric enhanced yellow fluorescent protein. Timelapse confocal imaging was performed to study the spatio-temporal movement of cSrc vesicles which formed from the plasma membrane. Analysis was facilitated by the use of various modules in the Volocity software package. The Visualization Module was used to view the entire timelapse dataset, which included multiple timepoints and z-sections. The [Quantitation] Module was used to track the intracellular movement of enlarged vesicles that originated from the plasma membrane. An easy-to-read spreadsheet was automatically generated which displayed the measurements for each timepoint that was tracked.