Custer C. Deocaris

Upregulation of mortalin/mthsp70/Grp75 contributes to human carcinogenesis

Mortalin, also known as mthsp70/GRP75/PBP74, interacts with the tumor suppressor protein p53 and inactivates its transcriptional activation and apoptotic functions. Here, we examined the level of mortalin expression in a large variety of tumor tissues, tumor‐derived and in vitro immortalized human cells. It was elevated in many human tumors, and in all of the tumor‐derived and in vitro immortalized cells. In human embryonic fibroblasts immortalized with an expression plasmid for hTERT, the telomerase catalytic subunit, with or without human papillomavirus E6 and E7 genes, we found that subclones with spontaneously increased mortalin expression levels became anchorage‐independent and acquired the ability to form tumors in nude mice. Furthermore, overexpression of mortalin was sufficient to increase the malignancy of breast carcinoma cells. The study demonstrates that upregulation of mortalin contributes significantly to tumorigenesis, and thus is a good candidate target for cancer therapy.

Three faces of mortalin : A housekeeper, guardian and killer

Mortalin was first cloned as a mortality factor that existed in the cytoplasmic fractions of normal, but not in immortal, mouse fibroblasts. A decade of efforts have expanded its persona from a house keeper protein involved in mitochondrial import, energy generation and chaperoning of misfolded proteins, to a guardian of stress that has multiple binding partners and to a killer protein that contributes to carcinogenesis on one hand and to old age disorders on the other. Being proved to be an attractive target for cancer therapy, it also warrants attention from the perspectives of management of old age diseases and healthy aging.

On the brotherhood of the mitochondrial chaperones mortalin and heat shock protein 60

Abstract The heat shock chaperones mortalin/mitochondrial heat shock protein 70 (mtHsp70) and Hsp60 are found in multiple subcellular sites and function in the folding and intracellular trafficking of many proteins. The chaperoning activity of these 2 proteins involves different structural and functional mechanisms. In spite of providing an excellent model for an evolutionarily conserved molecular “brotherhood,” their individual functions, although overlapping, are nonredundant. As they travel to various locations, both chaperones acquire different binding partners and exert a more divergent involvement in tumorigenesis, cellular senescence, and immunology. An understanding of their functional biology may lead to novel designing and development of therapeutic strategies for cancer and aging.

Threshold-like pattern of neuronal activation in the hypothalamus during treadmill running: establishment of a minimum running stress (MRS) rat model.

Despite the indication that the hypothalamo-pituitary-adrenal (HPA) axis is activated during treadmill running, there have not been any studies focusing on the relationship between exercise intensity and region-specific neural activities in hypothalamus. To address this, rats were subjected to 30 min of running, either at middle (supra-LT, 25 m min(-1)) or low speeds (sub-LT, 15 m min(-1)), and c-Fos-(+) cells were counted and compared with control rats. Significant increases in blood glucose and lactate levels, and plasma ACTH and osmolality levels were observed during supra-LT running. Only supra-LT running significantly increased c-Fos induction in various hypothalamic regions, namely, the medial preoptic area (MPO), periventricular nucleus (Pe), suprachiasmatic nucleus (SCN), supraoptic nucleus (SON), parvocellular division of the paraventricular nucleus (pPVN), anterior hypothalamic area (AH), arcuate nucleus (ARC) and posterior hypothalamic nucleus (PH). However, sub-LT caused no effect on c-Fos accumulation. This indicates that the hypothalamus responds uniquely to running in a threshold-like pattern distinct from the speed-dependent pattern previously reported for the medulla oblongata [Ohiwa et al., 2006a,b]. In addition, these results showed a physiologic basis for mild exercise useful for establishing our minimum running stress (MRS) rat model, or the running conditions that minimize the activation of the HPA axis.

Enhancing effect of cerebral blood volume by mild exercise in healthy young men: A near-infrared spectroscopy study

A mechanism by which exercise improves brain function may be attributed to increase in cerebral blood volume (CBV) with physical activity. However, the exact exercise intensity that influences CBV is still uncertain. To clarify this issue, 10 healthy young male participants were asked to perform a graded cycling exercise to the point of exhaustion while their prefrontal cortex CBVs are being monitored using near-infrared spectroscopy. Overall responsive cerebral oxygenation showed a non-linear pattern with three distinct phases. The CBV-threshold (CBVT), an event where rapid oxygenation takes place, occurred at approximately 42% of the V O2max. The CBVT preceded the lactate threshold (LT), which was at approximately 55% of the V O2max. The V O2max was not predictive of the CBVT in among the subjects. Our results indicate that oxygenation of the prefrontal cortex increases during graded cycling even at exercise intensities below the LT, suggesting the potential role of mild exercise in enhancing CBV.

Preincubation with the proteasome inhibitor MG-132 enhances proteasome activity via the Nrf2 transcription factor in aging human skin fibroblasts

Abstract:  Strategies that lead to the upregulation of the proteasome are known to elicit beneficial consequences to the organism by countering oxidative stress–associated disorders, such as protein conformational diseases, cancer, and aging. Mild treatment with proteasome inhibitors has been previously demonstrated to stimulate proteasome activity and cellular resistance against oxidative injury. However, the mechanism for this action has not been clearly defined. We examined the role of the nuclear factor‐E2‐related factor 2 (Nrf2) in fibroblasts, a key transactivator of the antioxidant response pathway, in the regulation of the proteasome by its inhibitor MG‐132. Here, we demonstrate that the stimulation of the proteasome by low levels of MG‐132 can be abrogated by small interfering RNAs (siRNAs) targeted against Nrf2. Consistently, cells that constitutively express Nrf2 exhibit elevated levels of proteasome activities. We further investigate how its beneficial effects, that is, proteasome stimulation, are manifested in young and replicative‐senescent cells. Our data underscore that manipulation of Nrf2 by the administration of pharmacologically low levels of proteasome inhibitors may prove to be an alternatively potent strategy for inducing long‐term protective effects against oxidative stress.

Functional Significance of Minor Structural and Expression Changes in Stress Chaperone Mortalin

Abstract:  Mortalin is one of the highly conserved heat‐shock chaperones. Some of the established features of mortalin include its various subcellular localizations, multiple binding partners, and differential subcellular distribution in normal and immortal cells. It inhibits nuclear translocation, transcriptional activation, and control of centrosome‐duplication functions of p53. It also functions as an adaptive protein in a variety of stress–response mechanisms and contributes to human carcinogenesis. Interestingly, minor alterations in its structure and level of expression may lead to drastic biological consequences (for example, Myelodysplastic syndrome and old age pathologies, such as Alzheimers and Parkinsons disease). Besides being validated as a reliable target for cancer therapy, mortalin also warrants attention from the perspectives of management of old‐age diseases and healthy aging.

Proteasomal Oscillation during Mild Heat Shock in Aging Human Skin Fibroblasts

Abstract:  Augmentation of proteasome machinery is emerging as a significant gerontomodulatory consequence of hormetic stimulation, such as mild heat stress. This study describes the phenomenon we term hormetic proteasomal oscillation, wherein mildly heat‐stressed human fibroblasts (41°C, 1 h) display an adaptation response pattern in proteasome activity. Remarkably, such response appears to be diverse in severely heat‐stressed or senescent fibroblasts. This proteasomal oscillation, as an innate cellular reaction to heat and aging, however, is independent of 20S proteasome protein levels and nuclear factor‐E2‐related factor 2 (Nrf2) transactivation.

Geroprotection by glycerol : Insights to its mechanisms and clinical potentials

Abstract:  Chaperones, particularly the heat‐shock proteins, are considered as key players in the maintenance of protein homeostasis and are associated with longevity and cellular immortalization. In this study, we investigated the geroprotective activity of the chemical chaperone glycerol. Glycerol showed significant chaperoning activity in refolding heat‐denatured luciferase in vivo and in protecting cells from heat stress‐induced cytotoxicity. This was accompanied by decrease in p53, an upregulation of a stress chaperone mortalin/mtHsp70, and an increase in proteasome activity in the presence of oxidative stress.

Glycerol stimulates innate chaperoning, proteasomal and stress-resistance functions: implications for geronto-manipulation

Aging is associated with accumulation of toxic intracellular and extracellular protein aggregates. Cells manage “aged” proteins by mobilizing their molecular chaperones or heat shock proteins that are also considered as determinants of lifespan in diverse species. In this study, we tested whether an exogenous addition of the non-toxic chemical chaperone ‘glycerol’ could elicit stress and geronto-protective activities. We found that glycerol enhanced chaperoning of heat-denatured proteins. In addition to stimulating proteasome activity, glycerol led to an increased expression of the stress chaperone ‘mortalin’ and decreased p53 function in human cells. Glycerol-fed worms exhibited thermo-tolerance and lower level of age-associated auto-fluorescence. Through the combined stimulation of the proteasome and chaperoning activities of mortalin, in particular, glycerol treatment resulted in increased survival and fitness against oxidative- and heat-stress. These results may have significant implications in the use of glycerol as a candidate geronto-modulator in development of practical interventions for “healthy aging”.