TY  - JOUR
A1  - Correia Santos, Sara
A1  - Bischler, Thorsten
A1  - Westermann, Alexander J.
A1  - Vogel, Jörg
T1  - MAPS integrates regulation of actin-targeting effector SteC into the virulence control network of Salmonella small RNA PinT
JF  - Cell Reports
N2  - A full understanding of the contribution of small RNAs (sRNAs) to bacterial virulence demands knowledge of their target suites under infection-relevant conditions. Here, we take an integrative approach to capturing targets of the Hfq-associated sRNA PinT, a known post-transcriptional timer of the two major virulence programs of Salmonella enterica. Using MS2 affinity purification and RNA sequencing (MAPS), we identify PinT ligands in bacteria under in vitro conditions mimicking specific stages of the infection cycle and in bacteria growing inside macrophages. This reveals PinT-mediated translational inhibition of the secreted effector kinase SteC, which had gone unnoticed in previous target searches. Using genetic, biochemical, and microscopic assays, we provide evidence for PinT-mediated repression of steC mRNA, eventually delaying actin rearrangements in infected host cells. Our findings support the role of PinT as a central post-transcriptional regulator in Salmonella virulence and illustrate the need for complementary methods to reveal the full target suites of sRNAs.
KW  - gene expression
KW  - nondocing RNA
KW  - chaperone HFQ
KW  - soluble-RNA
KW  - SEQ
KW  - interactome
KW  - repression
KW  - secretion
KW  - infection
KW  - biology
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-259134
VL  - 34
IS  - 5
ER  - 
TY  - JOUR
A1  - Eiring, Patrick
A1  - McLaughlin, Ryan
A1  - Matikonda, Siddharth S.
A1  - Han, Zhongying
A1  - Grabenhorst, Lennart
A1  - Helmerich, Dominic A.
A1  - Meub, Mara
A1  - Beliu, Gerti
A1  - Luciano, Michael
A1  - Bandi, Venu
A1  - Zijlstra, Niels
A1  - Shi, Zhen-Dan
A1  - Tarasov, Sergey G.
A1  - Swenson, Rolf
A1  - Tinnefeld, Philip
A1  - Glembockyte, Viktorija
A1  - Cordes, Thorben
A1  - Sauer, Markus
A1  - Schnermann, Martin J.
T1  - Targetable conformationally restricted cyanines enable photon-count-limited applications
JF  - Angewandte Chemie Internationale Edition
N2  - Cyanine dyes are exceptionally useful probes for a range of fluorescence-based applications, but their photon output can be limited by trans-to-cis photoisomerization. We recently demonstrated that appending a ring system to the pentamethine cyanine ring system improves the quantum yield and extends the fluorescence lifetime. Here, we report an optimized synthesis of persulfonated variants that enable efficient labeling of nucleic acids and proteins. We demonstrate that a bifunctional sulfonated tertiary amide significantly improves the optical properties of the resulting bioconjugates. These new conformationally restricted cyanines are compared to the parent cyanine derivatives in a range of contexts. These include their use in the plasmonic hotspot of a DNA-nanoantenna, in single-molecule Förster-resonance energy transfer (FRET) applications, far-red fluorescence-lifetime imaging microscopy (FLIM), and single-molecule localization microscopy (SMLM). These efforts define contexts in which eliminating cyanine isomerization provides meaningful benefits to imaging performance.
KW  - biology
KW  - super-resolution microscopy
KW  - conformational restriction
KW  - cyanine dyes
KW  - DNA nanotechnology
KW  - fluorescent dyes
KW  - single-molecule fluorescence spectroscopy
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-256559
VL  - 60
IS  - 51
ER  -