The conserved adenosine in helix 6 of Archaeoglobus fulgidus signal recognition particle RNA initiates SRP assembly

Jiaming Yin, Qiaojia Huang, Olga N. Pakhomova, Andrew P. Hinck, Christian W Zwieb

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The signal recognition particle (SRP) RNA helix 6 of archaea and eukaryotes is essential for the binding of protein SRP19 and the assembly of a functional complex. The conserved adenosine at the third position of the tetraloop of helix 6 (A149) is crucial for the binding of protein SRP19 in the mammalian SRP. Here we investigated the significance of the equivalent adenosine residue at position 159 (A159) of Archaeoglobus fulgidus SRP RNA. The A159 of A. fulgidus and A149 of human SRP RNA were changed to C, G or U, and fragments containing helix 6 or helices 6 and 8 were synthesized by run-off transcription with T7 RNA polymerase. The ability of recombinant A. fulgidus and human SRP19 to form ribonucleoprotein complexes was measured in vitro. The simultaneous presence of A149 and helix 8 is required for the high-affinity binding of SRP19 to the human SRP RNA. In contrast, A. fulgidus SRP19 binds to the SRP RNA fragments with high affinity irrespective of the nature of the nucleotide, demonstrating that A159 does not directly participate in protein binding. Instead, as indicated by the resistance of the wild-type A. fulgidus RNA towards digestion by RNase A, this residue allows the formation of a tightly folded RNA molecule. The high affinity between A. fulgidus SRP19 and RNA molecules that contain both helices 6 and 8 suggests that A159 is likely to initiate archaeal SRP assembly by forming a conserved tertiary RNA-RNA interaction.

Original languageEnglish (US)
Pages (from-to)269-275
Number of pages7
JournalArchaea
Volume1
Issue number4
StatePublished - Oct 2004

Fingerprint

Archaeoglobus
Archaeoglobus fulgidus
Signal Recognition Particle
adenosine
Adenosine
RNA
binding proteins
Carrier Proteins
protein
particle
ribonucleoproteins
Pancreatic Ribonuclease
Ribonucleoproteins
protein binding
Archaea
DNA-directed RNA polymerase
Eukaryota
Protein Binding
eukaryote
eukaryotic cells

Keywords

  • Protein-RNA interactions
  • Site-directed mutagenesis
  • Tetraloop

ASJC Scopus subject areas

  • Microbiology
  • Environmental Science(all)
  • Biotechnology

Cite this

Yin, J., Huang, Q., Pakhomova, O. N., Hinck, A. P., & Zwieb, C. W. (2004). The conserved adenosine in helix 6 of Archaeoglobus fulgidus signal recognition particle RNA initiates SRP assembly. Archaea, 1(4), 269-275.

The conserved adenosine in helix 6 of Archaeoglobus fulgidus signal recognition particle RNA initiates SRP assembly. / Yin, Jiaming; Huang, Qiaojia; Pakhomova, Olga N.; Hinck, Andrew P.; Zwieb, Christian W.

In: Archaea, Vol. 1, No. 4, 10.2004, p. 269-275.

Research output: Contribution to journalArticle

Yin, J, Huang, Q, Pakhomova, ON, Hinck, AP & Zwieb, CW 2004, 'The conserved adenosine in helix 6 of Archaeoglobus fulgidus signal recognition particle RNA initiates SRP assembly', Archaea, vol. 1, no. 4, pp. 269-275.
Yin, Jiaming ; Huang, Qiaojia ; Pakhomova, Olga N. ; Hinck, Andrew P. ; Zwieb, Christian W. / The conserved adenosine in helix 6 of Archaeoglobus fulgidus signal recognition particle RNA initiates SRP assembly. In: Archaea. 2004 ; Vol. 1, No. 4. pp. 269-275.
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