Determining the Sub-Cellular Localization of Proteins within Caenorhabditis elegans Body Wall Muscle

Meissner, Barbara; Rogalski, Teresa M.; Viveiros, Ryan; Warner, Adam; Plastino, Lorena; Lorch, Adam H.; Granger, Laure; Ségalat, Laurent; Moerman, Donald G.

doi:10.1371/journal.pone.0019937

Cited by 44 publications

(42 citation statements)

References 56 publications

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“…LGMD1D mutations were shown to result in stabilization and, therefore, increased levels of DNAJB6. While the amino acids associated with LGMD1D are not conserved in DNJ-24(Hsp40), DNJ-24(Hsp40) is enriched in muscle and shows the expected muscle and nuclear distribution pattern [49]. To address whether increased levels of this chaperones disrupted chaperone interactions in muscle cells, we examined the effects of muscle over-expression of dnj-24(Hsp40) (DNJ-24M) on synthetic motility defects induced by chaperone knock-down.…”

Section: Resultsmentioning

confidence: 99%

A Differentiation Transcription Factor Establishes Muscle-Specific Proteostasis in Caenorhabditis elegans

et al. 2016

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Safeguarding the proteome is central to the health of the cell. In multi-cellular organisms, the composition of the proteome, and by extension, protein-folding requirements, varies between cells. In agreement, chaperone network composition differs between tissues. Here, we ask how chaperone expression is regulated in a cell type-specific manner and whether cellular differentiation affects chaperone expression. Our bioinformatics analyses show that the myogenic transcription factor HLH-1 (MyoD) can bind to the promoters of chaperone genes expressed or required for the folding of muscle proteins. To test this experimentally, we employed HLH-1 myogenic potential to genetically modulate cellular differentiation of Caenorhabditis elegans embryonic cells by ectopically expressing HLH-1 in all cells of the embryo and monitoring chaperone expression. We found that HLH-1-dependent myogenic conversion specifically induced the expression of putative HLH-1-regulated chaperones in differentiating muscle cells. Moreover, disrupting the putative HLH-1-binding sites on ubiquitously expressed daf-21(Hsp90) and muscle-enriched hsp-12.2(sHsp) promoters abolished their myogenic-dependent expression. Disrupting HLH-1 function in muscle cells reduced the expression of putative HLH-1-regulated chaperones and compromised muscle proteostasis during and after embryogenesis. In turn, we found that modulating the expression of muscle chaperones disrupted the folding and assembly of muscle proteins and thus, myogenesis. Moreover, muscle-specific over-expression of the DNAJB6 homolog DNJ-24, a limb-girdle muscular dystrophy-associated chaperone, disrupted the muscle chaperone network and exposed synthetic motility defects. We propose that cellular differentiation could establish a proteostasis network dedicated to the folding and maintenance of the muscle proteome. Such cell-specific proteostasis networks can explain the selective vulnerability that many diseases of protein misfolding exhibit even when the misfolded protein is ubiquitously expressed.

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Section: Resultsmentioning

confidence: 99%

A Differentiation Transcription Factor Establishes Muscle-Specific Proteostasis in Caenorhabditis elegans

et al. 2016

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“…Expression of NRA-2 occurs largely in neurons, body wall muscle, and pharyngeal muscle, where it helps regulate acetylcholine receptor subunit composition, and RNAi knockdown of NRA-2 expression has been shown to sensitize C. elegans touch receptor neurons to Ca 2+ -mediated necrotic cell death (Kamat et al 2014). The next protein decreased in abundance MLP-1 is a LIM domain-containing cysteine-rich protein (CRP) expressed in a wide range of cell types in both larva and adults (including intestine, spermatheca, gonad sheath, hypodermis, body wall muscle, pharynx, and neurons) (McKay et al 2003; Meissner et al 2011). …”

Section: Resultsmentioning

confidence: 99%

Metabolome and proteome changes with aging in Caenorhabditis elegans

Copes

Edwards

Chaput

et al. 2015

Experimental Gerontology

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To expand the understanding of aging in the model organism Caenorhabditis elegans, global quantification of metabolite and protein levels in young and aged nematodes was performed using mass spectrometry. With age there was a decreased abundance of proteins functioning in transcription termination, mRNA degradation, mRNA stability, protein synthesis, and proteasomal function. Furthermore there was altered S-adenosyl methionine metabolism as well as a decreased abundance of the S-adenosyl methionine synthetase (SAMS-1) protein. Other aging-related changes included alterations in free fatty acid levels and composition, decreased levels of ribosomal proteins, decreased levels of NADP-dependent isocitrate dehydrogenase (IDH1), a shift in the cellular redox state, an increase in sorbitol content, alterations in free amino acid levels, and indications of altered muscle function and sarcoplasmic reticulum Ca2+ homeostasis. There were also decreases in pyrimidine and purine metabolite levels, most markedly nitrogenous bases. Supplementing the culture medium with cytidine (a pyrimidine nucleoside) or hypoxanthine (a purine base) increased lifespan slightly, suggesting that aging-induced alterations in ribonucleotide metabolism affect lifespan. An age-related increase in body size, lipotoxicity from ectopic yolk lipoprotein accumulation, a decline in NAD+ levels, and mitochondrial electron transport chain dysfunction may explain many of these changes. In addition, dietary restriction in aged worms resulting from sarcopenia of the pharyngeal pump likely decreases the abundance of SAMS-1, possibly leading to decreased phosphatidylcholine levels, larger lipid droplets, and ER and mitochondrial stress. The complementary use of proteomics and metabolomics yielded unique insights into the molecular processes altered with age in C. elegans.

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“…Selective RNAi KD of PEPCK-C or PYK-1, the PK isoform expressed in BDWM (39), in BDWM of post-fertile worms, reduced and extended lifespan, respectively ( Fig. 2G and supplemental Table S1).…”

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

confidence: 98%

“…C. elegans has pck-1, pck-2, and pck-3, three genes encoding PEPCK. PCK-1 localizes in cytosol (39) and thus is PEPCK-C. PCK-2 but not PCK-1 or PCK-3 has a predicted mitochondrial targeting sequence (data not shown), thus likely to be PEPCK-M. Global PEPCK-C knock-out (KO) in mice is lethal (40).…”

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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Determining the Sub-Cellular Localization of Proteins within Caenorhabditis elegans Body Wall Muscle

Cited by 44 publications

References 56 publications

A Differentiation Transcription Factor Establishes Muscle-Specific Proteostasis in Caenorhabditis elegans

A Differentiation Transcription Factor Establishes Muscle-Specific Proteostasis in Caenorhabditis elegans

Metabolome and proteome changes with aging in Caenorhabditis elegans

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

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