As guardians of the genome, RecQ helicases preserve our genetic coding and therefore protect our cells against aging. Likewise, Nuclear Pore Complexes (NPCs) maintain proper nuclear functionality through the transport of protein and mRNA. We hypothesize, from our preliminary work, that there exists a connection between RecQ helicases and specific proteins in the NPC that function together in maintenance of genomic longevity. Our goal is to gather quantitative data through targeted proteomics to prove this interdependence. Application of these methods will identify important RecQ helicases and Nups. We propose 1) to further test a panel of RecQ helicase knockout DT40 cells and mutant RecQ helicase fibroblasts for Nup protein and mRNA levels, 2) to analyze changes in nuclear transport by monitoring energy gradients and transport of NLS-GFP, 3) to obtain accurate measurements of NPC stoichiometry through targeted proteomics.
Aging involves the buildup of harmful alterations in the cell over time. One factor in aging is genome instability, which results from a decreased accuracy during replication. These errors lead to acceleration of the process. In our proposal, we focus on DNA damage repair complexes that are associated with the Nuclear Pore Complex to conceivably bringing double strand breaks to the nuclear periphery for repair. Impairment of these connections may be a factor in enhanced aging.
| Kosinski, Jan; Mosalaganti, Shyamal; von Appen, Alexander et al. (2016) Molecular architecture of the inner ring scaffold of the human nuclear pore complex. Science 352:363-5 |
| Hoelz, André; Glavy, Joseph S; Beck, Martin (2016) Toward the atomic structure of the nuclear pore complex: when top down meets bottom up. Nat Struct Mol Biol 23:624-30 |
| Castroagudin, Michelle R; Zhai, Yujia; Li, Zhi et al. (2016) Cyto-3D-print to attach mitotic cells. Cytotechnology 68:1641-5 |
| von Appen, Alexander; Kosinski, Jan; Sparks, Lenore et al. (2015) In situ structural analysis of the human nuclear pore complex. Nature 526:140-143 |
