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TitleThe box C/D sRNP dimeric architecture is conserved across domain Archaea.
Journal, issue, pagesRNA, Vol. 18, Issue 8, Page 1527-1540, Year 2012
Publish dateJun 29, 2012
AuthorsKathleen R Bower-Phipps / David W Taylor / Hong-Wei Wang / Susan J Baserga /
PubMed AbstractBox C/D small (nucleolar) ribonucleoproteins [s(no)RNPs] catalyze RNA-guided 2'-O-ribose methylation in two of the three domains of life. Recent structural studies have led to a controversy over ...Box C/D small (nucleolar) ribonucleoproteins [s(no)RNPs] catalyze RNA-guided 2'-O-ribose methylation in two of the three domains of life. Recent structural studies have led to a controversy over whether box C/D sRNPs functionally assemble as monomeric or dimeric macromolecules. The archaeal box C/D sRNP from Methanococcus jannaschii (Mj) has been shown by glycerol gradient sedimentation, gel filtration chromatography, native gel analysis, and single-particle electron microscopy (EM) to adopt a di-sRNP architecture, containing four copies of each box C/D core protein and two copies of the Mj sR8 sRNA. Subsequently, investigators used a two-stranded artificial guide sRNA, CD45, to assemble a box C/D sRNP from Sulfolobus solfataricus with a short RNA methylation substrate, yielding a crystal structure of a mono-sRNP. To more closely examine box C/D sRNP architecture, we investigate the role of the omnipresent sRNA loop as a structural determinant of sRNP assembly. We show through sRNA mutagenesis, native gel electrophoresis, and single-particle EM that a di-sRNP is the near exclusive architecture obtained when reconstituting box C/D sRNPs with natural or artificial sRNAs containing an internal loop. Our results span three distantly related archaeal species--Sulfolobus solfataricus, Pyrococcus abyssi, and Archaeoglobus fulgidus--indicating that the di-sRNP architecture is broadly conserved across the entire archaeal domain.
External linksRNA / PubMed:22753779 / PubMed Central
MethodsEM (single particle)
Resolution25.0 - 26.0 Å
Structure data

EMDB-5419:
Sulfolobus solfataricus methylation-guide sRNP
Method: EM (single particle) / Resolution: 26.0 Å

EMDB-5420:
Pyrococcus abyssi methylation-guide sRNP
Method: EM (single particle) / Resolution: 25.0 Å

EMDB-5421:
Archaeoglobus fulgidus methylation-guide sRNP
Method: EM (single particle) / Resolution: 26.0 Å

Source
  • Sulfolobus solfataricus (archaea)
  • Pyrococcus abyssi (archaea)
  • Archaeoglobus fulgidus (archaea)

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