Crystallization of the two-domain N-terminal fragment of the archaeal ribosomal protein L10(P0) in complex with a specif

  • PDF / 355,010 Bytes
  • 3 Pages / 612 x 792 pts (letter) Page_size
  • 34 Downloads / 162 Views

DOWNLOAD

REPORT


TAL GROWTH Dedicated to the memory of B.K. Vainshtein

Crystallization of the TwoDomain NTerminal Fragment of the Archaeal Ribosomal Protein L10(P0) in Complex with a Specific Fragment of 23S rRNA1 O. V. Kravchenko, I. V. Mitroshin, A. G. Gabdulkhakov, S. V. Nikonov, and M. B. Garber Institute of Protein Research RAS, Pushchino email: [email protected] Received February 28, 2011

Abstract—Lateral L12stalk (P1stalk in Archaea, P1/P2stalk in eukaryotes) is an obligatory morphological element of large ribosomal subunits in all organisms studied. This stalk is composed of the complex of ribo somal proteins L10(P0) and L12(P1) and interacts with 23S rRNA through the protein L10(P0). L12(P1)– stalk is involved in the formation of GTPase center of the ribosome and plays an important role in the ribo some interaction with translation factors. High mobility of this stalk puts obstacles in determination of its structure within the intact ribosome. Crystals of a twodomain Nterminal fragment of ribosomal protein L10(P0) from the archaeon Methanococcus jannaschii in complex with a specific fragment of rRNA from the same organism have been obtained. The crystals diffract Xrays at 3.2 Å resolution. DOI: 10.1134/S1063774511040109

INTRODUCTION In all organisms studied the large ribosomal subunit contains the functionally important lateral stalk called in Bacteria—L12stalk, in Archaea—P1stalk and in eukaryotes—P1/P2stalk [1]. This stalk is involved in formation of the GTPase center of the ribosome and plays an important role in the interaction of the ribo some with translation factors and in the control of the translation accuracy [2]. Isolated ribosomal proteins of this stalk form in solution a stable complex, which binds the large ribosomal RNA through the Ntermi nal domain of L10(P0) protein. Archaeal ribosomal proteins P0 and P1 differ from their bacterial counter parts L10 and L12 but are close by their primary struc ture to the eukaryal ribosomal proteins P0 and P1/P2. However, the structure of the RNAbinding domain of the proteins L10 and P0 is extremely conserved and therefore this domain is able to bind so conserved frag ment of the large ribosomal RNA from any organism. It was demonstrated that the archaeal and eukaryal complexes P0P1 are able to replace in vitro L10L12 complex in the bacterial ribosome, and the hybrid ribosome begins to interact with archaeal and eukary otic translational factors but not with own bacterial factors [3]. The structure of the ribosomal 50S subunit from the archaeon Haloarcula marismortui has been deter mined about 10 years ago at 2.4 Å resolution, but P1 stalk was absent in that model [4]. Later the structure 1The article was translated by the authors.

of the Nterminal RNAbinding domain of P0(L10) protein has been localized in this model of the 50S subunit at a low resolution level [5]. Recently this RNAbinding domain of L10 protein has been vizual ized on the electron density map of the bacterial 70S ribosome cocrystallized with elongation factor EFG [6]