Structural Basis for the Regulation of Biofilm Formation and Iron Uptake in A. baumannii by the Blue-Light-Using Photoreceptor, BlsA

Iva Chitrakar, James N. Iuliano, Yong Le He, Helena A. Woroniecka, Jinnette Tolentino Collado, Jinelle M. Wint, Stephen G. Walker, Peter J. Tonge, Jarrod B. French

Research output: Contribution to journalArticlepeer-review

Abstract

The opportunistic human pathogen, A. baumannii, senses and responds to light using the blue light sensing A (BlsA) photoreceptor protein. BlsA is a blue-light-using flavin adenine dinucleotide (BLUF) protein that is known to regulate a wide variety of cellular functions through interactions with different binding partners. Using immunoprecipitation of tagged BlsA in A. baumannii lysates, we observed a number of proteins that interact with BlsA, including several transcription factors. In addition to a known binding partner, the iron uptake regulator Fur, we identified the biofilm response regulator BfmR as a putative BlsA-binding partner. Using microscale thermophoresis, we determined that both BfmR and Fur bind to BlsA with nanomolar binding constants. To better understand how BlsA interacts with and regulates these transcription factors, we solved the X-ray crystal structures of BlsA in both a ground (dark) state and a photoactivated light state. Comparison of the light- A nd dark-state structures revealed that, upon photoactivation, the two α-helices comprising the variable domain of BlsA undergo a distinct conformational change. The flavin-binding site, however, remains largely unchanged from dark to light. These structures, along with docking studies of BlsA and Fur, reveal key mechanistic details about how BlsA propagates the photoactivation signal between protein domains and on to its binding partner. Taken together, our structural and biophysical data provide important insights into how BlsA controls signal transduction in A. baumannii and provides a likely mechanism for blue-light-dependent modulation of biofilm formation and iron uptake.

Original languageEnglish (US)
Pages (from-to)2592-2603
Number of pages12
JournalACS Infectious Diseases
Volume6
Issue number10
DOIs
StatePublished - Oct 9 2020

Bibliographical note

Funding Information:
J.N.I. and J.T.C. would like to acknowledge the Stony Brook Chemical Biology Training Program (T32GM092714) and the Stony Brook IMSD-MERGE Program (R25GM103962), respectively, for training support. This work was supported by the National Science Foundation through award numbers MCB-1750637 (J.B.F.) and MCB-1817837 (P.J.T.) and by the Research Corporation for Science Advancement through a Cottrell Scholar award to J.B.F. The content is solely the responsibility of the authors and does not necessarily represent the official views of any of the funding agencies.

Keywords

  • BLUF
  • BfmR
  • FUR
  • LOV
  • biofilms
  • photoreceptor

PubMed: MeSH publication types

  • Journal Article
  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

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