Regulation of transcription in hyperthermophilic archaea

A.B. Brinkman

Research output: Thesisinternal PhD, WU

Abstract

<p>The aim of the research presented here was to insight in the mechanisms by which transcription in hyperthermophilic archaea is regulated. To accomplish this, we have aimed (I) to identify transcriptional regulatory proteins from hyperthermophilic archaea, (II) to characterize these proteins, and (III) to determine how these proteins modulate the process of transcription initiation.</p><strong><p>Chapter 1</strong> describes the characteristics of the archaeal transcription machinery, and compiles transcription-related data that was obtained in the past two decades. The archaeal transcription machinery appears to be a simplified version of the eukaryal RNA polymerase II system, lacking various general transcription factors that are essential for eukaryal transcription initiation. However, archaeal genomes encode TFE, a homologue eukaryal TFIIE<FONT FACE="Symbol">a</font>transcription factor. Its stimulatory role in transcription is described in <strong>Chapter 2</strong> . <strong></p></strong><p>Although the archaeal transcription machinery is eukaryal-like, many genes encoding members of bacterial regulatory protein families can be found within archaeal genomes. Members of the Lrp family are most abundantly present in archaea and <strong>Chapter 3</strong> describes the properties of Lrp-like proteins. When this research project was started, fully sequenced archaeal genomes just became available . Only the gene encoding LrpA from <em>P. furiosus</em> had previously been identified in our laboratory and by others , and our initial strategy included the characterization of this protein, which is described in <strong>Chapter 4</strong> . LrpA was shown to negatively autoregulate its own transcription in a ligand-independent manner. The efficient production and purification of recombinant LrpA enabled crystallization of the protein and <strong>Chapter 5</strong> describes its resolved three-dimensional structure, which is the first structure of a member of the Lrp family. Subsequently, during the participation of our laboratory in the <em>S.solfataricus</em> P2 genome sequencing project, we were able to identify candidate regulatory genes in a more directed, bioinformatics-based approach, resulting in the identification and characterization of LysM and ChoR. LysM is another example of an archaeal Lrp-like protein, and in <strong>Chapter 6</strong> we have used the genomic context of LysM in the <em>S.solfataricus</em> genome to experimentally identify its physiological target and ligand. This study indicates for the first time that an Lrp-like protein may activate archaeal transcriptional.</p><p>Besides bacterial-like regulators, archaeal genomes encode unique archaeal-specific regulators that can be identified on the basis of a present DNA-binding domain. A putative regulator for <u>c</u> opper <u>ho</u> meostasis (ChoR) was identified in the <em>S.solfataricus</em> genome on the basis of a predicted HTH DNA-binding domain and a metal-binding domain. In <strong>Chapter 7</strong> it is demonstrated that ChoR is indeed a metal-responsive DNA-binding protein that is most likely involved in the repression of a heavy metal-efflux system.</p><strong><p>Chapter 8</strong> summarizes the data presented in this thesis, and adds some concluding remarks with respect to the implications of the work.
Original languageEnglish
QualificationDoctor of Philosophy
Awarding Institution
  • Wageningen University
Supervisors/Advisors
  • de Vos, W.M., Promotor
Award date6 Sep 2002
Place of PublicationS.l.
Print ISBNs9789058086730
Publication statusPublished - 2002

Keywords

  • thermophilic bacteria
  • transcription factors

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