2015
DOI: 10.1080/09168451.2015.1015950 View full text |Buy / Rent full text
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Abstract: Anabaena sensory rhodopsin transducer (ASRT) is believed to be a major player in the photo-signal transduction cascade, which is triggered by Anabaena sensory rhodopsin. Here, we characterized DNA binding activity of ASRT probed by using fluorescence correlation spectroscopy. We observed clear decrease of diffusion coefficient of DNA upon binding of ASRT. The dissociation constant, K(D), of ASRT to 20 bp-long DNA fragments lied in micro-molar range and varied moderately with DNA sequence. Our results suggest t… Show more

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“…To observe ASRT-induced DNA bending, excess amount of ASRT (100 μM) was put into the sample chamber and the changes in the FRET value of individual DNA was monitored. We used the salt condition of 50 mM NaCl which was used in the previous FCS and NMR studies for ASRT-DNA binding 28 , 30 . Note that 50 mM NaCl was too weak to induce spontaneous DNA bending (Figure S2 , upper left panel).…”
Section: Resultsmentioning
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“…To observe ASRT-induced DNA bending, excess amount of ASRT (100 μM) was put into the sample chamber and the changes in the FRET value of individual DNA was monitored. We used the salt condition of 50 mM NaCl which was used in the previous FCS and NMR studies for ASRT-DNA binding 28 , 30 . Note that 50 mM NaCl was too weak to induce spontaneous DNA bending (Figure S2 , upper left panel).…”
Section: Resultsmentioning
“…By using EMSA, it has been shown that multiple regions in the promoter region and transcription start sites of the phycoerythrocyanin gene have higher binding affinity with ASRT 29 . The FCS study 30 found a K D of ~ 10 μM between ASRT and 20 bp fragment of phycoerythrocyanin promotor region and reported a weak sequence dependent changes of the K D . Together, a hypothesis emerged: ASRT might induce changes in the structure of DNA, such as in the cases of CAP and AraC proteins, for gene regulation purpose.…”
Section: Discussionmentioning
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“…However, rather than pumping ions, it has been shown to regulate the expression of phycocyanin and phycoerythrocyanin, part of the phycobilisome light-harvesting complex, via a soluble 14-kDa cytoplasmic transducer (ASRT) (29)(30)(31)(32)(33)(34), found on a single operon (27). Unlike the haloarchaeal sensory rhodopsins, which interact with their cognate transmembrane transducers to mediate light-stimulated phototaxis (35,36), it has been proposed that ASR's photosignal is relayed to ASRT, which then translocates to the nucleus where it can bind specifically to the promoter regions of multiple genes (33,34). Other studies have shown evidence that ASR alone in E. coli can bind DNA and regulate gene activity upon photostimulation (31).…”
Section: Introductionmentioning