Article

Poly(ADP-ribose) is required for spindle assembly and structure

Department of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Nature (Impact Factor: 42.35). 01/2005; 432(7017):645-9. DOI: 10.1038/nature03061
Source: PubMed

ABSTRACT The mitotic spindle is typically thought of as an array of microtubules, microtubule-associated proteins and motors that self-organizes to align and segregate chromosomes. The major spindle components consist of proteins and DNA, the primary structural elements of the spindle. Other macromolecules including RNA and lipids also associate with spindles, but their spindle function, if any, is unknown. Poly(ADP-ribose) (PAR) is a large, branched, negatively charged polymeric macromolecule whose polymerization onto acceptor proteins is catalysed by a family of poly(ADP-ribose) polymerases (PARPs). Several PARPs localize to the spindle in vertebrate cells, suggesting that PARPs and/or PAR have a role in spindle function. Here we show that PAR is enriched in the spindle and is required for spindle function--PAR hydrolysis or perturbation leads to rapid disruption of spindle structure, and hydrolysis during spindle assembly blocks the formation of bipolar spindles. PAR exhibits localization dynamics that differ from known spindle proteins and are consistent with a low rate of turnover in the spindle. Thus, PAR is a non-proteinaceous, non-chromosomal component of the spindle required for bipolar spindle assembly and function.

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    • "As a result, scaffolds of protein-PAR::protein interactions are formed (reviewed in [18]). Such scaffolding property is observed in the recruitment of DNA repair proteins at the site of DNA damage [21], and for the assembly of spindle poles [22] and RNA organelles such as stress granules [14]. PARylation can be reversed mainly through two classes of degradation enzymes—one that can break the ribose–ribose bonds within the PAR chain and the other that breaks the covalent bonds between the proximal ADP-ribose units and the modified proteins. "
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    ABSTRACT: ADP-ribosylation refers to the addition of one or more ADP-ribose units onto protein substrates and this protein modification has been implicated in various cellular processes including DNA damage repair, RNA metabolism, transcription and cell cycle regulation. This review focuses on a compilation of large-scale proteomics studies that identify ADP-ribosylated proteins and their associated proteins by mass spectrometry using a variety of enrichment strategies. Some methods, such as the use of a poly(ADP-ribose)-specific antibody and boronate affinity chromatography and NAD+ analogues, have been employed for decades while others, such as the use of protein microarrays and recombinant proteins that bind ADP-ribose moieties (such as macrodomains), have only recently been developed. The advantages and disadvantages of each method and whether these methods are specific for identifying mono(ADP-ribosyl)ated and poly(ADP-ribosyl)ated proteins will be discussed. Lastly, since poly(ADP-ribose) is heterogeneous in length, it has been difficult to attain a mass signature associated with the modification sites. Several strategies on how to reduce polymer chain length heterogeneity for site identification will be reviewed.This article is protected by copyright. All rights reserved
    Proteomics 09/2014; 15(2-3). DOI:10.1002/pmic.201400217 · 3.97 Impact Factor
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    • "its assembly and function, although this was attributed to the enzymatic activity of another PARP family member, that is, tankyrase-1 [129] [130]. With regards to PARP1, it was shown that haploinsufficiency for PARP1 is related to centrosome duplication and chromosomal instability [131]. "
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    ABSTRACT: Aging is a multifactorial process that depends on diverse molecular and cellular mechanisms, such as genome maintenance and inflammation. The nuclear enzyme poly(ADP-ribose) polymerase 1 (PARP1), which catalyzes the synthesis of the biopolymer poly(ADP-ribose), exhibits an essential role in both processes. On the one hand, PARP1 serves as a genomic caretaker as it participates in chromatin remodelling, DNA repair, telomere maintenance, resolution of replicative stress, and cell cycle control. On the other hand, PARP1 acts as a mediator of inflammation due to its function as a regulator of NF-κB and other transcription factors and its potential to induce cell death. Consequently, PARP1 represents an interesting player in several aging mechanisms and is discussed as a longevity assurance factor on the one hand and an aging-promoting factor on the other hand. Here, we review the molecular mechanisms underlying the various roles of PARP1 in longevity and aging with special emphasis on cellular studies and we briefly discuss the results in the context of in vivo studies in mice and humans.
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    • "its assembly and function, although this was attributed to the enzymatic activity of another PARP family member, that is, tankyrase-1 [129] [130]. With regards to PARP1, it was shown that haploinsufficiency for PARP1 is related to centrosome duplication and chromosomal instability [131]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Aging is a multifactorial process that depends on diverse molecular and cellular mechanisms, such as genome maintenance and inflammation. The nuclear enzyme poly(ADP-ribose) polymerase 1 (PARP1), which catalyzes the synthesis of the biopolymer poly(ADP-ribose), exhibits an essential role in both processes. On the one hand, PARP1 serves as a genomic caretaker as it participates in chromatin remodelling, DNA repair, telomere maintenance, resolution of replicative stress, and cell cycle control. On the other hand, PARP1 acts as a mediator of inflammation due to its function as a regulator of NF-κB and other transcription factors and its potential to induce cell death. Consequently, PARP1 represents an interesting player in several aging mechanisms and is discussed as a longevity assurance factor on the one hand and an aging-promoting factor on the other hand. Here, we review the molecular mechanisms underlying the various roles of PARP1 in longevity and aging with special emphasis on cellular studies and we briefly discuss the results in the context of in vivo studies in mice and humans.
    Oxidative Medicine and Cellular Longevity 09/2012; 2012:321653. DOI:10.1155/2012/321653 · 3.36 Impact Factor
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