Hereditary Spastic Paraplegia Lab Report

The name of the homolog of my gene in C. elegans is act-3 and its gene id is 179533. It is similar to ACT-1, ACT-2, and ACT-4. Its locus is T04C12.4 and it is also known as ACTin family member (act-3).

The pET30a expression vector featuring T7 promoter, N/C-terminal His-tag sequence, T7 terminator and Lac operator was used for ligation of F protein gene at EcoRI restriction sites. The F gene-pET30a ligation was transformed into E.coli strain TOP10 competent cells and spread on Luria Bertani (LB) plates with kanamycin selection. The single colonies were picked and screened for positive cloning through EcoRI restriction digestion. The correct orientation of the gene was confirmed through restriction digestion of the PCR product with ScaI and EcoRV.

Spinal Muscular Atrophy, also referred to as SMA, is an incurable autosomal recessive disease caused by the mutation or deletion of the survival motor neuron 1(SMN1) gene located on chromosome five. This illness is the number one cause of infant death from a genetic disease. The disease is researched and studied heavily in the hopes to find an approved treatment and cure for the disease. With that being said, any discoveries on the disease are very important. In ones research of SMA, key features of the disease must be discussed in great detail. The cause of SMA, the symptoms, the diagnosis, the progression of the disease, impacts of being the victim of SMA , any possible treatments (if there are any), and the support that is provided for families of the patient (if support is provided or not) are all important aspects of the research. Prior research on the topic must also be considered when discussing the disease in order to understand SMA.

I aspire to be an orthopedic surgeon with a specialization in spinal surgery; our research has allowed me to familiarize myself with this region. The research I perform is centered on trying to understand the molecular mechanism by which mutations in a neuronal kinesin cause Hereditary Spastic Paraplegia. Though it is basic science research it has a transitional target. As a laboratory technician, I perform many of the complicated procedures myself and aid the undergraduate researchers in their projects whilst collaborating with our personal investigator Dr. Thomas Huckaba. Under his mentorship my capacity for critical reasoning has grown exponentially. As of now, my primary project is discovering the cargo that is being transported by Kinesin

actin “slid” past each other and neither filament changed in length. In 1957 Allan Huxley

MACF1 (microtubule cross linking factor 1) is large cytoskeletal linker protein (608 kda) that can closely linked with actin microfilament and microtubules (MTs). MACF1 gene is located on human chr 1p32, composed of about 110 exons and give rise to four major isoforms with slightly

Additionally, the Q555X nonsense mutation can reduce CaMKII and MAPK/ERK activity, which regulates the trafficking of synaptic vesicles and neurite outgrowth, and the missense mutation of A550T and T567A has been shown to impair synapsin targeting to nerve terminals (Fassio et al, 2011).

Myosin is a motor protein that uses energy from ATP hydrolysis to move along actin filaments. All myosins are composed of a 'tail' domain at their carboxy terminus and a globular 'head' domain at their amino terminus. The 'heads' contain sites for ATP binding. A complete round of ATP hydrolysis produces a single 'step' or movement of myosin along the actin

Moreover, further deletion mapping could be performed to narrow down the specific sequence within the third SIRPT of sacsin that binds JIP3.

It is generally accepted that resting CaMKIIβ bundles and stabilizes actin cytoskeleton. Transient activation of CaMKIIβ relaxes cytoskeleton, promotes actin polymerization and CaMKIIβ recruitment, and favors cytoskeleton growth. This model has been shown in both synaptogenesis (Okamoto et al., 2007) and OL maturation (Waggener et al., 2013). Based on this theory, we proposed that prolonged CaMKIIβ activation by NMDA-R-mediated Ca2+ influx leads to destabilization of actin cytoskeleton and membrane disintegration in mature OLs. Although our experiments unambiguously showed that NMDA-R activation results in CaMKIIβ activation, direct proof that links CaMKIIβ activation and membrane reduction are lacking. All the existing CaMKIIβ inhibitors

In this figure 1a, the DNase I performed the foot printing of K562 cells, they identify an individual nucleotide in the MTPN promoter. These promoters are bound by NRF1.In figure 1b, 8 defined

Introduction - Before examining the structural differences between TubZ and the rest of the Tubulin family, it will be necessary to give background on both. Specifically, what Tubulin is and the common characteristics of the Tubulin family, Tubulin’s role in the cell, what TubZ is, what it’s role is, and what similarities it shares with other members of the Tubulin family. A general layout of the paper will also be given at the end of this introduction. General layout would briefly describe the order of the sections to follow.

The whole network was analyzed by NetworkAnalyzer, which computes complex network parameters such as node degrees, average clustering coefficients, topological coefficients, and shortest path lengths. In the PPI network the nodes with high degree defined as hub proteins and the nodes with high betweenness defined as bottleneck proteins, which both play a fundamental role in the network [16]. Therefore, the degree and betweenness of each node are calculated, leading to identification of hub and bottleneck proteins (Table 5). The top 15 hub and bottleneck proteins are mainly those associated with ubiquitin, cytoskeleton and molecular chaperones. Hub and bottleneck proteins with their interacting proteins or chemicals were predicted from STICH web server and illustrated in Figure 3. More importantly, hub and bottleneck proteins within the network have been demonstrated to be potential therapeutic targets [17-19]. By submitting these hub and bottleneck proteins as drug targets in Drugbank database, potential drugs to treat MPS VII disease were predicted (Table 6).

So the first major step in the transfer of T strand is the formation of a long flexious appendage known as the T-pilus (Bulgakov, 2000) . The detection of presence of acetosyringone and other substances activates the expression of 11 genes in VirB operon which are responsible for the formation of the T-pilus. Other products of VirB genes like VirB6, VirB7 etc are used to transfer other subunits across the plasma memebrane.

Dysregulation of cytoskeletal genes has also been seen in MPS I mouse brains [4], MPS IIIB mouse brains [26] and MPS VII dog hearts [27]. Additionally, low density lipoprotein-related protein 1B (LRP1B), dynamin-1 (DMN1) and dynamin-related protein 1 (DNM1L), which are involved in the process of endocytosis, are downregulated. Vesicle associated membrane protein 2 (VAMP2) is involved in the targeting and/or fusion of transport vesicles to their target membrane [28]. These results further suggest impaired lysosomal membrane trafficking pathways in MPS VII.