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Single Molecule Study of RelA During...

Li, Wenting.

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Single Molecule Study of RelA During the Stringent Response in Live E. Coli Cells.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Single Molecule Study of RelA During the Stringent Response in Live E. Coli Cells./
Author:	Li, Wenting.
Description:	100 p.
Notes:	Source: Dissertation Abstracts International, Volume: 77-10(E), Section: B.
Contained By:	Dissertation Abstracts International77-10B(E).
Subject:	Analytical chemistry. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=10124176
ISBN:	9781339823027

Single Molecule Study of RelA During the Stringent Response in Live E. Coli Cells.
Li, Wenting.

Single Molecule Study of RelA During the Stringent Response in Live E. Coli Cells. - 100 p.

Source: Dissertation Abstracts International, Volume: 77-10(E), Section: B.

Thesis (Ph.D.)--The University of Wisconsin - Madison, 2016.

During amino acid starvation, bacterial cells rapidly synthesize the nucleotides (p)ppGpp, causing a massive re-programming of the transcriptional profile known as the stringent response. The (p)ppGpp synthase RelA is activated by ribosomes harboring an uncharged tRNA at the A site. It is unclear whether synthesis occurs while RelA is bound to the ribosome or free in the cytoplasm. We present a study of three E. coli strains, each expressing a different RelA-fluorescent protein (RelA-FP) construct: RelA-YFP, RelA-mEos2, and RelA-Dendra2. Single-molecule localization and tracking studies were carried out under normal growth conditions and during amino acid starvation. Study of three labeling schemes enabled us to assess potential problems with FP labeling of RelA. The diffusive trajectories and axial spatial distributions indicate that amino acid starvation induces net binding of all three RelA-FP constructs to 70S ribosomes. The data are most consistent with a model in which RelA synthesizes (p)ppGpp while bound to the 70S ribosome. We suggest a "short hopping time" model of RelA activity during starvation. Our results contradict an earlier study of RelA-Dendra2 diffusion that inferred off-ribosome synthesis of (p)ppGpp. The reasons for the discrepancy remain unclear.

ISBN: 9781339823027Subjects--Topical Terms:

3168300
Analytical chemistry.

Single Molecule Study of RelA During the Stringent Response in Live E. Coli Cells.
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100 1 $a Li, Wenting. $3 3276613
245 1 0 $a Single Molecule Study of RelA During the Stringent Response in Live E. Coli Cells.
300 $a 100 p.
500 $a Source: Dissertation Abstracts International, Volume: 77-10(E), Section: B.
500 $a Adviser: James C. Weisshaar.
502 $a Thesis (Ph.D.)--The University of Wisconsin - Madison, 2016.
520 $a During amino acid starvation, bacterial cells rapidly synthesize the nucleotides (p)ppGpp, causing a massive re-programming of the transcriptional profile known as the stringent response. The (p)ppGpp synthase RelA is activated by ribosomes harboring an uncharged tRNA at the A site. It is unclear whether synthesis occurs while RelA is bound to the ribosome or free in the cytoplasm. We present a study of three E. coli strains, each expressing a different RelA-fluorescent protein (RelA-FP) construct: RelA-YFP, RelA-mEos2, and RelA-Dendra2. Single-molecule localization and tracking studies were carried out under normal growth conditions and during amino acid starvation. Study of three labeling schemes enabled us to assess potential problems with FP labeling of RelA. The diffusive trajectories and axial spatial distributions indicate that amino acid starvation induces net binding of all three RelA-FP constructs to 70S ribosomes. The data are most consistent with a model in which RelA synthesizes (p)ppGpp while bound to the 70S ribosome. We suggest a "short hopping time" model of RelA activity during starvation. Our results contradict an earlier study of RelA-Dendra2 diffusion that inferred off-ribosome synthesis of (p)ppGpp. The reasons for the discrepancy remain unclear.
520 $a Here is the paper I wrote on my dissertation work.
590 $a School code: 0262.
650 4 $a Analytical chemistry. $3 3168300
650 4 $a Biophysics. $3 518360
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710 2 $a The University of Wisconsin - Madison. $b Chemistry. $3 2049906
773 0 $t Dissertation Abstracts International $g 77-10B(E).
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792 $a 2016
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856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=10124176