Proteomics and electron microscopic characterization of the unusual mitochondrial ribosome-related 45S complex in Leishmania tarentolae.
A novel type of ribonucleoprotein (RNP) complex has been described from the kinetoplast-mitochondria of Leishmania tarentolae. The complex, termed the 45S SSU*, contains the 9S small subunit rRNA but does not contain the 12S large subunit rRNA. This complex is the most stable and abundant mitochondrial RNP complex present in Leishmania. As shown by tandem mass spectrometry, the complex contains at least 39 polypeptides with a combined molecular mass of almost 2.1 MDa. These components include several homologs of small subunit ribosomal proteins (S5, S6, S8, S9, S11, S15, S16, S17, S18, MRPS29); however, most of the polypeptides present are unique. Only a few of them show recognizable motifs, such as protein-protein (coiled-coil, Rhodanese) or protein-RNA (pentatricopeptide repeat) interaction domains. A cryo-electron microscopy examination of the 45S SSU* fraction reveals that 27% of particles represent SSU homodimers arranged in a head-to-tail orientation, while the majority of particles are clearly different and show an asymmetric bilobed morphology. Multiple classes of two-dimensional averages were derived for the asymmetrical particles, probably reflecting random orientations of the particles and difficulties in correlating these views with the known projections of ribosomal complexes. One class of the two-dimensional averages shows a SSU moiety attached to a protein mass or masses in a monosome-like appearance. The combined mass spectrometry and electron microscopy data thus indicate that the majority 45S SSU* particles represents a heterodimeric complex in which the SSU of the Leishmania mitochondrial ribosome is associated with an additional protein mass. The biological role of these particles is not known.
Maslov
Dmitri A
DA
Department of Biology, University of California, Riverside, CA 92521, USA. maslov@ucr.edu
Spremulli
Linda L
LL
Sharma
Manjuli R
MR
Bhargava
Kalpana
K
Grasso
Domenick
D
Falick
Arnold M
AM
Agrawal
Rajendra K
RK
Parker
Carol E
CE
Simpson
Larry
L
D000818:Animals; D020285:Cryoelectron Microscopy; D007891:Leishmania / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics; D015800:Protozoan Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D012335:RNA, Ribosomal / Q000737:chemistry; D012261:Ribonucleoproteins / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D012269:Ribosomal Proteins / Q000737:chemistry; D053719:Tandem Mass Spectrometry
D024101:Mitochondrial Proteins; D021122:Protein Subunits; D015800:Protozoan Proteins; D012335:RNA, Ribosomal; D012261:Ribonucleoproteins; D012269:Ribosomal Proteins