34 research outputs found
Mechanistic Insight into the Reactivation of BCAII Enzyme from Denatured and Molten Globule States by Eukaryotic Ribosomes and Domain V rRNAs
In all life forms, decoding of messenger-RNA into polypeptide chain is accomplished by the ribosome. Several protein chaperones are known to bind at the exit of ribosomal tunnel to ensure proper folding of the nascent chain by inhibiting their premature folding in the
densely crowded environment of the cell. However, accumulating evidence suggests that ribosome may play a chaperone role in protein folding events in vitro. Ribosome-mediated folding of denatured proteins by prokaryotic ribosomes has been studied extensively. The
RNA-assisted chaperone activity of the prokaryotic ribosome has been attributed to the domain V, a span of 23S rRNA at the intersubunit side of the large subunit encompassing
the Peptidyl Transferase Centre. Evidently, this functional property of ribosome is unrelated to the nascent chain protein folding at the exit of the ribosomal tunnel. Here, we seek to scrutinize whether this unique function is conserved in a primitive kinetoplastid group of eukaryotic species Leishmania donovani where the ribosome structure possesses distinct additional features and appears markedly different compared to other higher eukaryotic
ribosomes. Bovine Carbonic Anhydrase II (BCAII) enzyme was considered as the model protein. Our results manifest that domain V of the large subunit rRNA of Leishmania ribosomes
preserves chaperone activity suggesting that ribosome-mediated protein folding is, indeed, a conserved phenomenon. Further, we aimed to investigate the mechanism underpinning the ribosome-assisted protein reactivation process. Interestingly, the surface plasmon resonance binding analyses exhibit that rRNA guides productive folding by directly interacting with molten globule-like states of the protein. In contrast, native protein shows no
notable affinity to the rRNA. Thus, our study not only confirms conserved, RNA-mediated chaperoning role of ribosome but also provides crucial insight into the mechanism of the process
Mechanisms of Hearing Loss after Blast Injury to the Ear
Given the frequent use of improvised explosive devices (IEDs) around the world, the study of traumatic blast injuries is of
increasing interest. The ear is the most common organ affected by blast injury because it is the bodyï¾’s most sensitive
pressure transducer. We fabricated a blast chamber to re-create blast profiles similar to that of IEDs and used it to develop a
reproducible mouse model to study blast-induced hearing loss. The tympanic membrane was perforated in all mice after
blast exposure and found to heal spontaneously. Micro-computed tomography demonstrated no evidence for middle ear or
otic capsule injuries; however, the healed tympanic membrane was thickened. Auditory brainstem response and distortion
product otoacoustic emission threshold shifts were found to be correlated with blast intensity. As well, these threshold
shifts were larger than those found in control mice that underwent surgical perforation of their tympanic membranes,
indicating cochlear trauma. Histological studies one week and three months after the blast demonstrated no disruption or
damage to the intra-cochlear membranes. However, there was loss of outer hair cells (OHCs) within the basal turn of the
cochlea and decreased spiral ganglion neurons (SGNs) and afferent nerve synapses. Using our mouse model that
recapitulates human IED exposure, our results identify that the mechanisms underlying blast-induced hearing loss does not
include gross membranous rupture as is commonly believed. Instead, there is both OHC and SGN loss that produce auditory
dysfunction
Environmental Enrichment Preceding Early Adulthood Methylphenidate Treatment Leads to Long Term Increase of Corticosterone and Testosterone in the Rat
Attention-deficit/hyperactivity disorder (ADD/ADHD) has been emerging as a world-wide psychiatric disorder. There appears to be an increasing rate of stimulant drug abuse, specifically methylphenidate (MPH) which is the most common treatment for ADHD, among individuals who do not meet the criteria for ADHD and particularly for cognitive enhancement among university students. However, the long term effects of exposure to MPH are unknown. Thus, in light of a developmental approach in humans, we aimed to test the effects of adolescence exposure to enriched environment (EE) followed by MPH administration during early adulthood, on reactions to stress in adulthood. Specifically, at approximate adolescence [post natal days (PND) 30–60] rats were reared in EE and were treated with MPH during early adulthood (PND 60–90). Adult (PND 90–92) rats were exposed to mild stress and starting at PND 110, the behavioral and endocrine effects of the combined drug and environmental conditions were assessed. Following adolescence EE, long term exposure to MPH led to decreased locomotor activity and increased sucrose preference. EE had a beneficial effect on PPI (attentive abilities), which was impaired by long term exposure to MPH. Finally, the interaction between EE and, exposure to MPH led to long-term elevated corticosterone and testosterone levels. In view of the marked increase in MPH consumption over the past decade, vigilance is crucial in order to prevent potential drug abuse and its long term detrimental consequences