Site-Specific Turn-On Fluorescent Labeling of DNA-Interacting Protein Using Oligodeoxynucleotides That Modify Lysines To Produce 5,6-Dimethoxy 3-Methyleneisoindolin-1-one

ABSTRACT Several vectors that facilitate stable fluorescent labeling of Burkholderia pseudomallei and Burkholderia thailandensis were constructed. These vectors combined the effectiveness of the mini-Tn7 site-specific transposition system with fluorescent proteins optimized for Burkholderia spp., enabling bacterial tracking during cellular infection.

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DNA-binding regulates site-specific ubiquitination of IRF-1

Biochemical Journal ◽

10.1042/bj20121076 ◽

2013 ◽

Vol 449 (3) ◽

pp. 707-717 ◽

Cited By ~ 16

Author(s):

Vivien Landré ◽

Emmanuelle Pion ◽

Vikram Narayan ◽

Dimitris P. Xirodimas ◽

Kathryn L. Ball

Keyword(s):

Dna Binding ◽

E3 Ligase ◽

Binding Domain ◽

Dna Binding Domain ◽

Docking Site ◽

Interacting Protein ◽

Site Specific ◽

E3 Ligases ◽

Intrinsically Disordered ◽

C Terminus

Understanding the determinants for site-specific ubiquitination by E3 ligase components of the ubiquitin machinery is proving to be a challenge. In the present study we investigate the role of an E3 ligase docking site (Mf2 domain) in an intrinsically disordered domain of IRF-1 [IFN (interferon) regulatory factor-1], a short-lived IFNγ-regulated transcription factor, in ubiquitination of the protein. Ubiquitin modification of full-length IRF-1 by E3 ligases such as CHIP [C-terminus of the Hsc (heat-shock cognate) 70-interacting protein] and MDM2 (murine double minute 2), which dock to the Mf2 domain, was specific for lysine residues found predominantly in loop structures that extend from the DNA-binding domain, whereas no modification was detected in the more conformationally flexible C-terminal half of the protein. The E3 docking site was not available when IRF-1 was in its DNA-bound conformation and cognate DNA-binding sequences strongly suppressed ubiquitination, highlighting a strict relationship between ligase binding and site-specific modification at residues in the DNA-binding domain. Hyperubiquitination of a non-DNA-binding mutant supports a mechanism where an active DNA-bound pool of IRF-1 is protected from polyubiquitination and degradation.

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Site-Specific Fluorescent Labeling Approaches for Naringenin, an Essential Flavonone in Plant Nitrogen-Fixation Signaling Pathways

The Journal of Organic Chemistry ◽

10.1021/jo8014165 ◽

2008 ◽

Vol 73 (21) ◽

pp. 8279-8285 ◽

Cited By ~ 4

Author(s):

Lei Chen ◽

Feng-Qing Li ◽

Bi-He Hou ◽

Guo-Fan Hong ◽

Zhu-Jun Yao

Keyword(s):

Nitrogen Fixation ◽

Signaling Pathways ◽

Fluorescent Labeling ◽

Site Specific ◽

Plant Nitrogen

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Fluorescent turn-on probes for wash-free mRNA imaging via covalent site-specific enzymatic labeling

Chemical Science ◽

10.1039/c7sc03150e ◽

2017 ◽

Vol 8 (10) ◽

pp. 7169-7173 ◽

Cited By ~ 11

Author(s):

Cun Yu Zhou ◽

Seth C. Alexander ◽

Neal K. Devaraj

Keyword(s):

Cellular Regulation ◽

Site Specific ◽

Turn On ◽

The Many ◽

And Function

Investigating the many roles RNA plays in cellular regulation and function has increased demand for tools to explore RNA tracking and localization within cells.

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