As proteins age they are more likely to acquire molecular damage. To combat the functional decline of the proteome, most cellular proteins are rapidly turned over. In this way, potentially impaired polypeptides are constantly replaced with newly translated copies. Proteins with slower rates of turnover are therefore at greater risk of suffering a deleterious modification. Recently, long-lived proteins (LLPs) were discovered in post-mitotic cells of the rat central nervous system. Strikingly, these LLPs are associated with chromatin, the nuclear envelope, and the plasma membrane, all of which are cellular compartments that coordinate a myriad of regulatory functions. LLPs are therefore constantly exposed to potentially harmful metabolites. It is unknown how the functional integrity of these proteins is maintained over such a long time period, and whether the longevity of these proteins plays a specific role in long-lived cells. Thus, proposed experiments will test the specific hypothesis that LLPs regulate biological processes over extremely long time frames and that their functional decline drives cellular and organismal aging. This research proposal is designed to study the biology of LLPs, primarily focusing on nucleoporins and histones, and how they relate to post-mitotic tissue function. First, quantitative mass spectrometry and multi- isotope imaging mass spectrometry will be used to measure cell, organelle, and protein turnover rates in post-mitotic tissues in rats. Second, biochemical, cell biological, and genome-wide approaches will be used to determine the molecular properties of long-lived nucleoporins and histones. Efforts will be directed toward identifying and characterizing protein-repair pathways and protective post-translational modifications that are responsible for LLP longevity. In addition, it will be determined whether long-lived nucleoporins and histones serve as molecular timers to regulate nuclear pore function and gene expression, respectively. Finally, LLPs will be depleted in post-mitotic cells using a novel protein extraction and degradation system to determine how LLP impairment affects cellular function and aging. These analyses will focus on aberrant intracellular trafficking and gene regulation. Proposed studies promise to establish novel links between protein longevity and aging, and provide new molecular targets for understanding and potentially treating age-associated degenerative disorders.
We recently discovered that neurons within the adult rat brain contain long-lived proteins that somehow escape protein degradation pathways designed to maintain a functional proteome. We plan to decipher the mechanisms by which the functional integrity of these proteins is protected over long periods of time, and determine whether their eventual functional decline contributes to age-related pathologies in post-mitotic tissues. Proposed experiments promise to reveal new biological factors and pathways that influence protein homeostasis, and provide new avenues for studying the 'normal' aging process and for developing therapies against age-related disease.
|McCloskey, Asako; Ibarra, Arkaitz; Hetzer, Martin W (2018) Tpr regulates the total number of nuclear pore complexes per cell nucleus. Genes Dev 32:1321-1331|
|Toda, Tomohisa; Hsu, Jonathan Y; Linker, Sara B et al. (2017) Nup153 Interacts with Sox2 to Enable Bimodal Gene Regulation and Maintenance of Neural Progenitor Cells. Cell Stem Cell 21:618-634.e7|
|Franks, Tobias M; McCloskey, Asako; Shokirev, Maxim Nikolaievich et al. (2017) Nup98 recruits the Wdr82-Set1A/COMPASS complex to promoters to regulate H3K4 trimethylation in hematopoietic progenitor cells. Genes Dev 31:2222-2234|
|Buchwalter, Abigail; Hetzer, Martin W (2017) Nucleolar expansion and elevated protein translation in premature aging. Nat Commun 8:328|
|van de Ven, Robert A H; de Groot, Jolien S; Park, Danielle et al. (2016) p120-catenin prevents multinucleation through control of MKLP1-dependent RhoA activity during cytokinesis. Nat Commun 7:13874|
|Hatch, Emily M; Hetzer, Martin W (2016) Nuclear envelope rupture is induced by actin-based nucleus confinement. J Cell Biol 215:27-36|
|Franks, Tobias M; Benner, Chris; Narvaiza, Iñigo et al. (2016) Evolution of a transcriptional regulator from a transmembrane nucleoporin. Genes Dev 30:1155-71|
|Ibarra, Arkaitz; Benner, Chris; Tyagi, Swati et al. (2016) Nucleoporin-mediated regulation of cell identity genes. Genes Dev 30:2253-2258|