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Real-time tracking reveals catalytic roles for the two DNA binding sites of Rad51.

ABSTRACT: During homologous recombination, Rad51 forms a nucleoprotein filament on single-stranded DNA to promote DNA strand exchange. This filament binds to double-stranded DNA (dsDNA), searches for homology, and promotes transfer of the complementary strand, producing a new heteroduplex. Strand exchange proceeds via two distinct three-strand intermediates, C1 and C2. C1 contains the intact donor dsDNA whereas C2 contains newly formed heteroduplex DNA. Here, we show that the conserved DNA binding motifs, loop 1 (L1) and loop 2 (L2) in site I of Rad51, play distinct roles in this process. L1 is involved in formation of the C1 complex whereas L2 mediates the C1-C2 transition, producing the heteroduplex. Another DNA binding motif, site II, serves as the DNA entry position for initial Rad51 filament formation, as well as for donor dsDNA incorporation. Our study provides a comprehensive molecular model for the catalytic process of strand exchange mediated by eukaryotic RecA-family recombinases.

SUBMITTER: Ito K

PROVIDER: S-EPMC7289862 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature

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Real-time tracking reveals catalytic roles for the two DNA binding sites of Rad51.

Ito Kentaro K Murayama Yasuto Y Kurokawa Yumiko Y Kanamaru Shuji S Kokabu Yuichi Y Maki Takahisa T Mikawa Tsutomu T Argunhan Bilge B Tsubouchi Hideo H Ikeguchi Mitsunori M Takahashi Masayuki M Iwasaki Hiroshi H

Nature communications 20200611 1

During homologous recombination, Rad51 forms a nucleoprotein filament on single-stranded DNA to promote DNA strand exchange. This filament binds to double-stranded DNA (dsDNA), searches for homology, and promotes transfer of the complementary strand, producing a new heteroduplex. Strand exchange proceeds via two distinct three-strand intermediates, C1 and C2. C1 contains the intact donor dsDNA whereas C2 contains newly formed heteroduplex DNA. Here, we show that the conserved DNA binding motifs, ...[more]

PMID: 32528002

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Project description:Airborne pollen monitoring has been an arduous task, making ecological applications and allergy management virtually disconnected from everyday practice. Over the last decade, intensive research has been conducted worldwide to automate this task and to obtain real-time measurements. The aim of this study was to evaluate such an automated biomonitoring system vs. the conventional 'gold-standard' Hirst-type technique, attempting to assess which may more accurately provide the genuine exposure to airborne pollen. Airborne pollen was monitored in Augsburg since 2015 with two different methods, a novel automatic Bio-Aerosol Analyser, and with the conventional 7-day recording Hirst-type volumetric trap, in two different sites. The reliability, performance, accuracy, and comparability of the BAA500 Pollen Monitor (PoMo) vs. the conventional device were investigated, by use of approximately 2.5 million particles sampled during the study period. The observations made by the automated PoMo showed an average accuracy of approximately 85%. However, it also exhibited reliability problems, with information gaps within the main pollen season of between 17 to 19 days. The PoMo automated algorithm had identification issues, mainly confusing the taxa of Populus, Salix and Tilia. Hirst-type measurements consistently exhibited lower pollen abundances (median of annual pollen integral: 2080), however, seasonal traits were more comparable, with the PoMo pollen season starting slightly later (median: 3 days), peaking later (median: 5 days) but also ending later (median: 14 days). Daily pollen concentrations reported by Hirst-type traps vs. PoMo were significantly, but not closely, correlated (r = 0.53-0.55), even after manual classification. Automatic pollen monitoring has already shown signs of efficiency and accuracy, despite its young age; here it is suggested that automatic pollen monitoring systems may be more effective in capturing a larger proportion of the airborne pollen diversity. Even though reliability issues still exist, we expect that this new generation of automated bioaerosol monitoring will eventually change the aerobiological era, as known for almost 70 years now.

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Real-time tracking reveals catalytic roles for the two DNA binding sites of Rad51.

Publications

Real-time tracking reveals catalytic roles for the two DNA binding sites of Rad51.

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