Monitoring Newly Synthesized Proteins over the Adult Life Span of <i>Caenorhabditis elegans</i>

Vukoti, Krishna; Yu, Xiaokun; Sheng, Quanhu; Saha, Sudipto; Feng, Zhaoyang; Hsu, Ao Lin; Miyagi, Masaru

doi:10.1021/acs.jproteome.5b00021

Cited by 23 publications

(29 citation statements)

References 39 publications

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“…In day 5 adult worms, average THPLH is much longer, which is in line with observations by Vukoti et al (2015). This study used a pulsed SILAC-based approach similar to ours to study protein turnover in wild-type worms during adulthood and found that protein turnover drastically dropped after day 5.…”

Section: Discussionsupporting

confidence: 87%

“…For determination of THPLH by pulsed SILAC, the adult worms have the benefit that small differences in synchronization have less effect than they do in developing worms and that the total protein concentration per worm stays largely the same over the course of the experiment. However, total label incorporation is lower (Vukoti et al, 2015), which makes estimation of THPLH less reliable. Worms were pulse labeled for 6, 24, and 48 hr and harvested.…”

Section: Protein Turnover Rates In Aging Wormsmentioning

confidence: 99%

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Proteome-wide Changes in Protein Turnover Rates in C. elegans Models of Longevity and Age-Related Disease

et al. 2016

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The balance between protein synthesis and protein breakdown is a major determinant of protein homeostasis, and loss of protein homeostasis is one of the hallmarks of aging. Here we describe pulsed SILAC-based experiments to estimate proteome-wide turnover rates of individual proteins. We applied this method to determine protein turnover rates in Caenorhabditis elegans models of longevity and Parkinson's disease, using both developing and adult animals. Whereas protein turnover in developing, long-lived daf-2(e1370) worms is about 30% slower than in controls, the opposite was observed in day 5 adult worms, in which protein turnover in the daf-2(e1370) mutant is twice as fast as in controls. In the Parkinson's model, protein turnover is reduced proportionally over the entire proteome, suggesting that the protein homeostasis network has a strong ability to adapt. The findings shed light on the relationship between protein turnover and healthy aging.

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Section: Discussionsupporting

confidence: 87%

Section: Protein Turnover Rates In Aging Wormsmentioning

confidence: 99%

Proteome-wide Changes in Protein Turnover Rates in C. elegans Models of Longevity and Age-Related Disease

et al. 2016

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“…Decline in PEPCK activity after the reproductive peak also occurred in glp-1 mutants, validating that it is not a result of loss of eggs. The reciprocal changes in PEPCK-C and PK with age also cannot be an effect of aging-associated loss of protein homeostasis (41), which predicts declines in both. We conclude that aging involves a programmed metabolic event that includes reciprocal changes in PEPCK-C and PK.…”

Section: Aging Involves a Progressive Decline In Pepck-c And A Reciprmentioning

confidence: 99%

Reciprocal Changes in Phosphoenolpyruvate Carboxykinase and Pyruvate Kinase with Age Are a Determinant of Aging in Caenorhabditis elegans

Yuan¹,

Hakimi²,

Kao³

et al. 2016

Journal of Biological Chemistry

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Aging involves progressive loss of cellular function and integrity, presumably caused by accumulated stochastic damage to cells. Alterations in energy metabolism contribute to aging, but how energy metabolism changes with age, how these changes affect aging, and whether they can be modified to modulate aging remain unclear. In locomotory muscle of post-fertile Caenorhabditis elegans, we identified a progressive decrease in cytosolic phosphoenolpyruvate carboxykinase (PEPCK-C), a longevity-associated metabolic enzyme, and a reciprocal increase in glycolytic pyruvate kinase (PK) that were necessary and sufficient to limit lifespan. Decline in PEPCK-C with age also led to loss of cellular function and integrity including muscle activity, and cellular senescence. Genetic and pharmacologic interventions of PEPCK-C, muscle activity, and AMPK signaling demonstrate that declines in PEPCK-C and muscle function with age interacted to limit reproductive life and lifespan via disrupted energy homeostasis. Quantifications of metabolic flux show that reciprocal changes in PEPCK-C and PK with age shunted energy metabolism toward glycolysis, reducing mitochondrial bioenergetics. Last, calorie restriction countered changes in PEPCK-C and PK with age to elicit antiaging effects via TOR inhibition. Thus, a programmed metabolic event involving PEPCK-C and PK is a determinant of aging that can be modified to modulate aging.Aging is characterized by the progressive decline in cellular function and integrity that leads to disease vulnerability and eventually death of organisms (1). The leading proposed cause of decline in cellular function and integrity with age is the accumulation of stochastic damage of molecules and organelles by reactive molecules, such as reactive oxygen species (ROS). Whether ROS are detrimental to organisms and whether ROS limit lifespan, however, are in debate (2).Energy metabolism supplies ATP for cellular function and maintenance. Alterations in energy metabolism are linked to the aging process and aging-associated diseases (3). In model organisms, environmental and genetic factors that change energy metabolism, such as calorie restriction (CR) (4), inhibition of target of rapamycin (TOR) (5), and 5Ј AMP kinase (AMPK) (6) are determinants of longevity. A large body of aging research has been focusing on the signaling of CR, TOR inhibition, and AMPK in regulating longevity. The exact alterations in energy metabolism that occur with age, how these changes impact aging, and whether they can be modified to modulate aging are understudied and remain poorly understood, largely due to the intrinsic complexity of energy metabolism, and the indirect impact of these longevity paradigms on energy metabolism. This impedes the understanding of aging mechanisms and the development of mechanism-based strategies to modulate aging.A key regulation of energy metabolism at the cellular level is the reciprocal changes of PK and PEPCK-C (7). Whether this regulation of cellular energy metabolism contributes to organismal aging is...

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“…In recent years, proteomics analyses have shown alterations in protein abundance and its expression control in aged organisms, providing insights into the association of individual proteins with the aging process (David et al 2010;Mao et al 2010;Walther & Mann 2011;Reis-Rodrigues et al 2012;Janssens et al 2015;Vukoti et al 2015;Walther et al 2015;Narayan et al 2016). However, proteomics studies on asymmetric inheritance for individual damaged or old-age proteins likely to influence cellular aging have been limited.…”

Section: Introductionmentioning

confidence: 99%

Proteomics analysis for asymmetric inheritance of preexisting proteins between mother and daughter cells in budding yeast

et al. 2017

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In budding yeast, a mother cell can produce a finite number of daughter cells over its life. The accumulation of a variety of types of damaged components has an impact on the aging process. Asymmetrical inheritance during cell division causes these aberrant intracellular constituents to be retained in mother cells and prevents them from segregating to daughter cells. However, the understanding of asymmetrical inheritance of individual proteins that are damaged or old age, and their relevance to the aging process, has been limited. The aim of this study is to propose a proteomics strategy for asymmetrical inheritance of preexisting proteins between mother and daughter cells. During synchronous culture for one generation, newly synthesized proteins were labeled with stable isotope amino acids to discriminate preexisting proteins originally expressed in mother cells, followed by separation of mother and daughter cells using a conventional method based on biotin labeling. Isotope incorporation ratios for individual proteins were quantified using mass spectrometry. We successfully identified 21 proteins whose preexisting versions were asymmetrically inherited in mother cells, including plasma membrane transporter involved in the aging process and organelle-anchoring proteins related to the stress response to misfolded proteins. Thus, our approach would be useful for making catalog of asymmetrically inherited proteins.

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Monitoring Newly Synthesized Proteins over the Adult Life Span of Caenorhabditis elegans

Cited by 23 publications

References 39 publications

Proteome-wide Changes in Protein Turnover Rates in C. elegans Models of Longevity and Age-Related Disease

Proteome-wide Changes in Protein Turnover Rates in C. elegans Models of Longevity and Age-Related Disease

Reciprocal Changes in Phosphoenolpyruvate Carboxykinase and Pyruvate Kinase with Age Are a Determinant of Aging in Caenorhabditis elegans

Proteomics analysis for asymmetric inheritance of preexisting proteins between mother and daughter cells in budding yeast

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