Title

Determination and dissection of dna-binding specificity for the thermus thermophilus hb8 transcriptional regulator tthb099

Department

Chemistry and Biochemistry

Document Type

Article

Publication Date

11-1-2020

Abstract

© 2020 by the authors. Licensee MDPI, Basel, Switzerland. Transcription factors (TFs) have been extensively researched in certain well-studied organisms, but far less so in others. Following the whole-genome sequencing of a new organism, TFs are typically identified through their homology with related proteins in other organisms. However, recent findings demonstrate that structurally similar TFs from distantly related bacteria are not usually evolutionary orthologs. Here we explore TTHB099, a cAMP receptor protein (CRP)-family TF from the extremophile Thermus thermophilus HB8. Using the in vitro iterative selection method Restriction Endonuclease Protection, Selection and Amplification (REPSA), we identified the preferred DNA-binding motif for TTHB099, 5′ –TGT(A/g)NBSYRSVN(T/c)ACA–3′, and mapped potential binding sites and regulated genes within the T. thermophilus HB8 genome. Comparisons with expression profile data in TTHB099-deficient and wild type strains suggested that, unlike E. coli CRP (CRPEc ), TTHB099 does not have a simple regulatory mechanism. However, we hypothesize that TTHB099 can be a dual-regulator similar to CRPEc .

Journal Title

International Journal of Molecular Sciences

Journal ISSN

16616596

Volume

21

Issue

21

First Page

1

Last Page

18

Digital Object Identifier (DOI)

10.3390/ijms21217929

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