Nuclear Routing Networks Span between Nuclear Pore Complexes and Genomic DNA to Guide Nucleoplasmic Trafficking of Biomolecules
|Marek Malecki1* and Bianca Malecki2|
|1University of Wisconsin, Madison, WI, USA|
|2Phoenix Biomolecular Engineering Foundation, San Francisco, CA, USA|
|*Corresponding Author :||Marek Malecki, MD, PhD
University of Wisconsin
Madison, WI, USA
E-mail: [email protected]
|Received September 12, 2012; Accepted October 17, 2012; Published October 19, 2012|
|Citation: Malecki M, Malecki B (2012) Nuclear Routing Networks Span between Nuclear Pore Complexes and Genomic DNA to Guide Nucleoplasmic Trafficking of Biomolecules. J Fert In Vitro 2:112. doi:10.4172/2165-7491.1000112|
|Copyright: © 2012 Malecki M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.|
In health and disease, biomolecules, which are involved in gene expression, recombination, or reprogramming have to traffic through the nucleoplasm, between Nuclear Pore Complexes (NPCs) and genomic DNA (gDNA). This trafficking is guided by the recently revealed Nuclear Routing Networks (NRNs).
In this study, we aimed to investigate, if the NRNs have established associations with the genomic DNA in situ and if the NRNs have capabilities to bind the DNA de novo. Moreover, we aimed to study further, if nucleoplasmic trafficking of the histones, rRNA, and transgenes’ vectors, between the NPCs and gDNA, is guided by the NRNs.
We used Xenopus laevis oocytes as the model system. We engineered the transgenes’ DNA vectors equipped with the SV40 LTA Nuclear Localization Signals (NLS) and/or HIV Rev Nuclear Export Signals (NES). We purified histones, 5S rRNA, and gDNA. We rendered all these molecules superparamagnetic and fluorescent for detection with Nuclear Magnetic Resonance (NMR), Total Reflection X-Ray Fluorescence (TXRF), Energy Dispersive X-Ray Spectroscopy (EDXS), and Electron Energy Loss Spectroscopy (EELS).
The NRNs span between the NPCs and genomic DNA. They form firm bonds with the gDNA in situ. After
complete digestion of the nucleic acids with the RNases and DNases, the newly added DNA - modified with the dNTP analogs, bonds firmly to the NRNs. Moreover, the NRNs guide the trafficking of the DNA transgenes’ vectorsmodified with the SV40 LTA NLS, following their import into the nuclei through the NPCs. The pathway is identical to that of histones. The NRNs also guide the trafficking of the DNA transgenes’ vectors, modified with the HIV Rev NES, to the NPCs, followed by their export out of the nuclei. Ribosomal RNAs follow the same pathway.
To summarize, the NRNs are the structures connecting the NPCs and the gDNA. They guide the trafficking of the biomolecules between the NPCs and gDNA.