Research Associate Professor
Department of Pharmaceutical Sciences
College of Pharmacy
University of Nebraska Medical Center
986025 Nebraska Medical Center
Omaha, NE 68198-6025
My associates and I use a combination of bulk-phase biochemical and single-molecule biophysical approaches to understand the molecular basis of stalled DNA replication fork rescue, an essential process required to maintain genome stability. Our experimental work is concerned with the function and regulation of the complexes that control fork rescue, with studies focused primarily on the role of the single-strand DNA binding protein (SSB) and several recombination enzymes, using Escherichia coli as the model organism. The modern bulk-phase, biochemical approaches we use include spectroscopic (UV and Visible) and equilibrium binding. The single-molecule studies employ what we call “visual biochemistry”, a technique I pioneered that combines optical tweezers, microfluidics, and high-resolution fluorescence microscopy. To achieve single base-pair resolution, we also use magnetic tweezers, a complementary single-molecule microscopy technique. To observe fork rescue in living cells, we have developed a high-speed super-resolution microscopy technique based on structured illumination. We collaborate with researchers in the Department of Pharmaceutical Sciences, the Eppley Institute for Cancer Research, and colleagues in Australia, the UK, and China.
To learn about the details of available projects, students and researchers are welcome to visit the Bianco laboratory.
A. Fork rescue and the single-strand DNA binding protein
- Bianco, P. R. (2021) The mechanism of action of the SSB interactome reveals it is the first OB-fold family of genome guardians in prokaryotes. Protein Science, in press.
- Tan, H. Y., and Bianco, P.R. (2021) SSB facilitates fork substrate discrimination by PriA. ACS Omega, in press
- Bianco, P. R., and Lu, Y. (2021) Single molecule studies of stalled DNA replication fork rescue in E.coli. Nucleic Acids Research, 49(8):4220-4238
- Sun, Z., Wang, Y., Bianco, P.R. and Lyubchenko, Y.L. (2021) Nanoscale Dynamics of the PriA Helicase at Stalled DNA Replication Forks. J Phys Chem B. 2021 May 6;125(17):4299-4307
- Lu, Y., and Bianco, P. R. (2021) High yield purification of exceptional-quality DNA substrates suitable for single-molecule studies. J Biological Methods 8, 1-9
- Wang, Y., Sun, Z., Bianco, P. R., and Lyubchenko, Y. L. (2020) Atomic force microscopy-based characterization of the interaction of PriA helicase with stalled DNA replication forks. J Biol Chem 295, 6043-6052
- Ding, W., Tan, H. Y., Zhang, J. X., Wilczek, L. A., Hsieh, K. R., Mulkin, J. A., and Bianco, P. R. (2020) The mechanism of Single strand binding protein-RecG binding: Implications for SSB interactome function. Protein Sci 29, 1211-1227
- Bianco, P. R. (2020) DNA Helicase-SSB Interactions Critical to the Regression and Restart of Stalled DNA Replication forks in Escherichia coli. Genes (Basel) 11
- Zhao, T., Liu, Y., Wang, Z., He, R., Xiang Zhang, J., Xu, F., Lei, M., Deci, M. B., Nguyen, J., and Bianco, P. R. (2019) Super-resolution imaging reveals changes in Escherichia coli SSB localization in response to DNA damage. Genes Cells 24, 814-826
- Liu, X., Seet, J. X., Shi, Y., and Bianco, P. R. (2019) Rep and UvrD Antagonize One Another at Stalled Replication Forks and This Is Exacerbated by SSB. ACS Omega 4, 5180-5196
- Sun, Z., Hashemi, M., Warren, G., Bianco, P. R., and Lyubchenko, Y. L. (2018) Dynamics of the Interaction of RecG Protein with Stalled Replication Forks. Biochemistry 57, 1967-1976
- Bianco, P. R. (2018) RecA protein. Encyclopedia of Life Sciences Tan, H. Y., Wilczek, L. A., Pottinger, S., Manosas, M., Yu, C., Nguyenduc, T., and Bianco, P. R. (2017) The intrinsically disordered linker of E. coli SSB is critical for the release from single-stranded DNA. Protein Sci 26, 700-717
Bianco, P. R., Pottinger, S., Tan, H. Y., Nguyenduc, T., Rex, K., and Varshney, U. (2017) The IDL of E. coli SSB links ssDNA and protein binding by mediating protein-protein interactions. Protein Sci 26, 227-241
Bianco, P. R., and Lyubchenko, Y. L. (2017) SSB and the RecG DNA helicase: an intimate association to rescue a stalled replication fork. Protein Sci 26, 638-649
Bianco, P. R. (2017) The tale of SSB. Progress in biophysics and molecular biology 127, 111-118
B. Microscope development
- Zhao, T., Z. Wang, T. Chen, M. Lei, and P.R. Bianco. (2021) Recent Advances in High-Speed Structured Illumination Microscopy. Frontiers in Physics, 2021. in press.
- Zhao, T., U. Hao, Z. Wang, Y. Liang, K. Fweng, C. He, M., X. Yun, P.R. Bianco, Y. Sun, and M. Lei. (2021) Multi-color structured illumination microscopy for live cell imaging based on the enhanced image recombination transform algorithm. Biomedical Optics Express, 12(6) 3474-3484.
- He, M., Y. Liang, P.R. Bianco, Z. Wang, X. Yun, Y. Cai, K. Feng, and M. Lei. (2021) Trapping performance of holographic optical tweezers generated with different hologram algorithms. AIP Advances 11:035130.
- Dan, D., Wang, Z., Zhou, X., Lei, M., Zhao, T., Qian, J., Yu, X., Yan, S., Min, J., Bianco, P.R., Yao, B. (2021). Rapid image reconstruction of structured illumination microscopy directly in the spatial domain. IEEE Photonics Journal, 13(1): p. 1-11.
- Ren, F., Z. Wang, J. Qian, Y. Liang, S. Dang, Y. Cai, P.R. Bianco, B. Yao, and M. Lei. (2019) Multi-view object topography measurement with optical sectioning structured illumination microscopy. Appl Opt. 58(23): p. 6288-6294.
- Liang, Y., Y. Cai, Z. Wang, M. Lei, Z. Cao, Y. Wang, M. Li, S. Yan, P.R. Bianco, and B. Yao. (2018) Aberration correction in holographic optical tweezers using a high-order optical vortex. Appl Opt. 57(13): p. 3618-3623.
- Z. Wang, Y. Cai, Y. Liang, X. Zhou, S. Yan, D. Dan, P.R. Bianco, M. Lei, and B. Yao. (2017) Single shot three-dimensional fluorescent microscopy with a spatially rotating point spread function. Biomedical Optics Express. 8(12): p. 5493-5506.
C. Book Chapters
- Yu, C. and Bianco, P.R. (2021) In Oliveira, M. T. (ed.), Methods Molecular Biology, Single Stranded DNA Binding Proteins, Vol. 2281.
- Lyubchenko, Y.L., Sun, Z. and Bianco, P.R. (2017) In Lyubchenko, Y. L. (ed.), Introduction to single molecule biophysics. Chapter 4. Magnetic Tweezers. CRC Press, Taylor & Francis Group LLC.