6 research outputs found
The immunochemical cross-reactivity between cytoplasmic and mitochondrial mammalian lysyl-tRNA synthetases
Animal and fungal cells (in contrast to prokaryotes) contain two distinct sets of related aminoacyl-tRNA synthetases (aaRSs) encoded by nuclear genes and functioning in cytosol and mitochondria. The structural differences between mitochondrial and cytoplasmic enzymes may reflect the functional adaptation to fulfil mitochondrial processes in addition to protein synthesis. Mitochondrial import of nuclearencoded tRNAs has been described in yeast, plants and protozoans but it has not been observed in mammalian cells. Ifs established that mitochondrial lysyl-tRNA synthetase (MSK) plays a prominent role in the transport of tRNA into yeast mitochondria for complementation o f mitochondrial tRNAs genes mutations. We tried to identify MSK homologues in mammalian cells with the help of monospecific antibodies against pre-MSK by ELISA and Western-blot analysis. We have identified cross-reactive proteins in mitochondrial and cytoplasmic fractions of mammalian cell lysates. These data, together with the results of cross-aminoacylation on mitochondrial and cytoplasmic tRNAs, suggest the presence of common antigenic determinants in the mitochondrial and cytoplasmic lysyl-tRNA synthetases from higher animals.ΠΠ»ΡΡΠΈΠ½ΠΈ Π΅ΡΠΊΠ°ΡΡΠΎΡ Π½Π° Π²ΡΠ΄ΠΌΡΠ½Ρ Π²ΡΠ΄ ΠΏΡΠΎΠΊΠ°ΡΡΠΎΡ ΠΌΡΡΡΡΡΡ Π΄Π²Ρ ΡΡΠ·Π½Ρ Π³ΡΡΠΏΠΈ Π°ΠΌΡΠ½ΠΎΠ°ΡΠΈΠ»-ΡΠ ΠΠ ΡΠΈΠ½ΡΠ΅ΡΠ°Π·, ΡΠΊΡ ΠΊΠΎΠ΄ΡΡΡΡΡΡ ΡΠ΄Π΅ΡΠ½ΠΈΠΌ Π³Π΅Π½ΠΎΠΌΠΎΠΌ ΡΠ° ΡΡΠ½ΠΊΡΡΠΎΠ½ΡΡΡΡ Π² ΡΠΈΡΠΎΠ·ΠΎΠ»Ρ Ρ ΠΌΡΡΠΎΡ
ΠΎΠ½Π΄ΡΡΡΡ
. Π‘ΡΡΡΠΊΒΡΡΡΠ½Ρ Π²ΡΠ΄ΠΌΡΠ½Π½ΠΎΡΡΡ ΠΌΡΠΆ ΡΠ΅ΡΠΌΠ΅Π½ΡΠ°ΠΌΠΈ ΠΌΡΡΠΎΡ
ΠΎΠ½Π΄ΡΡΠΉ Ρ ΡΠΈΡΠΎΠΏΠ»Π°Π·ΒΠΌΠΈ ΠΌΠΎΠΆΡΡΡ Π±ΡΡΠΈ Π²ΡΠ΄ΠΎΠ±ΡΠ°ΠΆΠ΅Π½Π½ΡΠΌ ΡΡΠ½ΠΊΡΡΠΎΠ½Π°Π»ΡΠ½ΠΎΡ Π°Π΄Π°ΠΏΡΠ°ΡΡΡ Π΄ΠΎ ΠΏΡΠΎΡΠ΅ΡΡΠ², ΡΠΊΡ Π²ΡΠ΄Π±ΡΠ²Π°ΡΡΡΡΡ Π² ΠΌΡΡΠΎΡ
ΠΎΠ½Π΄ΡΡΡΡ
, ΠΊΡΡΠΌ ΡΡΠ°ΡΡΡ Π² Π±ΡΠΎΡΠΈΠ½ΡΠ΅Π·Ρ Π±ΡΠ»ΠΊΠ°. ΠΠΌΠΏΠΎΡΡ ΡΠΈΡΠΎΠ·ΠΎΠ»ΡΠ½ΠΈΡ
ΡΠ ΠΠ Ρ ΠΌΡΡΠΎΡ
ΠΎΠ½Π΄ΡΡΡ ΠΎΠΏΠΈΡΠ°Π½ΠΎ Π΄Π»Ρ Π΄ΡΡΠΆΠ΄ΠΆΡΠ², ΡΠΎΡΠ»ΠΈΠ½ Ρ Π½Π°ΠΉΠΏΡΠΎΡΡΡΠΈΡΠΈΡ
, ΠΎΠ΄Π½Π°ΠΊ Π²ΡΠ½ Π½Π΅ ΡΠΏΠΎΡΡΠ΅ΡΡΠ³Π°Π²ΡΡ Π² ΠΊΠ»ΡΡΠΈΠ½Π°Ρ
ΡΡΠ°Π²ΡΡΠ². ΠΠΈΡΠ²Π»Π΅Π½ΠΎ, ΡΠΎ ΠΌΡΡΠΎΡ
ΠΎΠ½Π΄ΡΡΠ°Π»ΡΠ½Π° Π»ΡΠ·ΠΈΠ»-ΡΠ ΠΠ ΡΠΈΠ½ΡΠ΅ΡΠ°Π·Π° (MSK) Π²ΡΠ΄ΡΠ³ΡΠ°Ρ ΠΏΡΠΎΠ²ΡΠ΄Π½Ρ ΡΠΎΠ»Ρ Ρ ΡΡΠ°Π½ΡΠΏΠΎΡΡΡ ΡΠ ΠΠ Ρ ΠΌΡΡΠΎΡ
ΠΎΠ½Π΄ΡΡΡ Π΄ΡΡΠΆΠ΄ΠΆΡΠ² Π΄Π»Ρ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΌΠ΅Π½ΒΡΠ°ΡΡΡ ΠΌΡΡΠ°ΡΡΠΉ ΠΌΡΡΠΎΡ
ΠΎΠ½Π΄ΡΡΠ°Π»ΡΠ½ΠΈΡ
Π³Π΅Π½ΡΠ² ΡΠ ΠΠ ΠΠ° Π΄ΠΎΠΏΠΎΠΌΠΎΠ³ΠΎΡ ΠΌΠΎΠ½ΠΎΡΠΏΠ΅ΡΠΈΡΡΡΠ½ΠΈΡ
Π°Π½ΡΠΈΡΡΠ» ΠΏΡΠΎΡΠΈ npe-MSK ΠΌΠΈ Π·ΡΠΎΠ±ΠΈΠ»ΠΈ ΡΠΏΡΠΎΠ±Ρ ΡΠ΄Π΅Π½ΡΠΈΡΡΠΊΡΠ²Π°ΡΠΈ Π³ΠΎΠΌΠΎΠ»ΠΎΠ³ΠΈ MSK Ρ ΠΊΠ»ΡΡΠΈΠ½Π°Ρ
ΡΡΠ°Π²ΡΡΠ² ΠΌΠ΅ΡΠΎΠ΄Π°ΠΌΠΈ ELISA Ρ ΠΠ΅ΡΡΠ΅ΡΠ½-Π±Π»ΠΎΡΠΈΠ½Π³Π°. Π ΡΠΈΡΠΎΠΏΠ»Π°Π·ΠΌΠ°ΡΠΈΡΠ½ΠΈΡ
Ρ ΠΌΡΡΠΎΒΡ
ΠΎΠ½Π΄ΡΡΠ°Π»ΡΠ½ΠΈΡ
ΡΡΠ°ΠΊΡΡΡΡ
Π»ΡΠ·Π°ΡΡΠ² ΠΊΠ»ΡΡΠΈΠ½ ΡΡΠ°Π²ΡΡΠ² Π½Π°ΠΌ Π²Π΄Π°Π»ΠΎΡΡ Π²ΠΈΡΠ²ΠΈΡΠΈ Π±ΡΠ»ΠΊΠΈ, ΡΠΊΡ ΠΌΠ°ΡΡΡ ΡΠΌΡΠ½ΠΎΠ»ΠΎΠ³ΡΡΠ½ΠΈΠΉ ΠΏΠ΅ΡΠ΅Ρ
ΡΠ΅ΡΡ Π· MSK Π Π°Π·ΠΎΠΌ Π· ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΠ°ΠΌΠΈ ΠΏΠ΅ΡΠ΅Ρ
ΡΠ΅ΡΠ½ΠΎΠ³ΠΎ Π°ΠΌΡΠ½ΠΎΠ°ΡΠΈΠ»ΡΠ²Π°Π½Π½Ρ ΡΡ Π΄Π°Π½Ρ Π΄Π°ΡΡΡ ΠΏΡΠ΄ΡΡΠ°Π²Ρ Π΄Π»Ρ ΠΏΡΠΈΠΏΡΡΠ΅Π½Π½Ρ ΡΠΎΠ΄ΠΎ Π½Π°ΡΠ²Π½ΠΎΡΡΡ ΡΠΏΡΠ»ΡΠ½ΠΈΡ
Π°Π½ΡΠΈΠ³Π΅Π½Π½ΠΈΡ
Π΄Π΅ΡΠ΅ΡΠΌΡΠ½Π°Π½Ρ Ρ ΠΌΡΡΠΎΡ
ΠΎΠ½Π΄ΡΡΠ°Π»ΡΠ½ΠΈΡ
Ρ ΡΠΈΡΠΎΠΏΠ»Π°Π·ΠΌΠ°ΒΡΠΈΡΠ½ΠΈΡ
Π»ΡΠ·ΠΈΠ»-ΡΠ ΠΠ ΡΠΈΠ½ΡΠ΅ΡΠ°Π· ΡΡΠ°Π²ΡΡΠ².ΠΠ»Π΅ΡΠΊΠΈ ΡΡΠΊΠ°ΡΠΈΠΎΡ (Π² ΠΎΡΠ»ΠΈΡΠΈΠ΅ ΠΎΡ ΠΊΠ»Π΅ΡΠΎΠΊ ΠΏΡΠΎΠΊΠ°ΡΠΈΠΎΡ) ΡΠΎΠ΄Π΅ΡΠΆΠ°Ρ Π΄Π²Π΅ ΡΠ°Π·Π»ΠΈΡΠ½ΡΠ΅ Π³ΡΡΠΏΠΏΡ Π°ΠΌΠΈΠ½ΠΎΠ°ΡΠΈΠ»-ΡΠ ΠΠ ΡΠΈΠ½ΡΠ΅ΡΠ°Π·, ΠΊΠΎΠ΄ΠΈΡΡΠ΅ΒΠΌΡΡ
ΡΠ΄Π΅ΡΠ½ΡΠΌ Π³Π΅Π½ΠΎΠΌΠΎΠΌ ΠΈ ΡΡΠ½ΠΊΡΠΈΠΎΠ½ΠΈΡΡΡΡΠΈΡ
Π² Ρ ΠΈ ΡΠΎ Π·ΠΎΠ»Π΅ ΠΈ ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΡΡ
. Π‘ΡΡΡΠΊΡΡΡΠ½ΡΠ΅ ΠΎΡΠ»ΠΈΡΠΈΡ ΠΌΠ΅ΠΆΠ΄Ρ ΡΠ΅ΡΠΌΠ΅Π½ΡΠ°ΠΌΠΈ ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠΉ ΠΈ ΡΠΈΡΠΎΠΏΠ»Π°Π·ΠΌΡ ΠΌΠΎΠ³ΡΡ Π±ΡΡΡ ΠΎΡΡΠ°ΠΆΠ΅Π½ΠΈΠ΅ΠΌ ΡΡΠ½ΠΊΡΠΈΒΠΎΠ½Π°Π»ΡΠ½ΠΎΠΉ Π°Π΄Π°ΠΏΡΠ°ΡΠΈΠΈ ΠΊ ΠΏΡΠΎΡΠ΅ΡΡΠ°ΠΌ, ΠΏΡΠΎΠΈΡΡ
ΠΎΠ΄ΡΡΠΈΠΌ Π² ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΒΡΠΈΡΡ
ΠΏΠΎΠΌΠΈΠΌΠΎ ΡΡΠ°ΡΡΠΈΡ Π² Π±ΠΈΠΎΡΠΈΠ½ΡΠ΅Π·Π΅ Π±Π΅Π»ΠΊΠ° ΠΠΌΠΏΠΎΡΡ ΡΠΈΡΠΎΠ·ΠΎΠ»ΡΠ½ΡΡ
ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠΈ ΠΎΠΏΠΈΡΠ°Π½ Ρ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ, ΡΠ°ΡΡΠ΅Π½ΠΈΠΉ ΠΈ ΠΏΡΠΎΡΡΠ΅ΠΉΡΠΈΡ
, ΠΎΠ΄Π½Π°ΠΊΠΎ Π½Π΅ Π½Π°Π±Π»ΡΠ΄Π°Π»ΡΡ Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ
ΠΌΠ»Π΅ΠΊΠΎΠΏΠΈΡΠ°ΡΒΡΠΈΡ
. Π£ΡΡΠ°Π½ΠΎΠ²Π»Π΅Π½ΠΎ, ΡΡΠΎ ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½Π°Ρ Π»ΠΈΠ·ΠΈΠ»-ΡΠ ΠΠ ΡΠΈΠ½ΒΡΠ΅ΡΠ°Π·Π° (MSK) ΠΈΠ³ΡΠ°Π΅Ρ Π²Π΅Π΄ΡΡΡΡ ΡΠΎΠ»Ρ Π² ΡΡΠ°Π½ΡΠΏΠΎΡΡΠ΅ ΡΠ ΠΠ Π² ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠΈ Π΄ΡΠΎΠΆΠΆΠ΅ΠΉ Π΄Π»Ρ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΌΠ΅Π½ΡΠ°ΡΠΈΠΈ ΠΌΡΡΠ°ΡΠΈΠΉ ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΡΡ
Π³Π΅Π½ΠΎΠ² ΡΠ ΠΠ. Π‘ ΠΏΠΎΠΌΠΎΡΡΡ ΠΌΠΎΠ½ΠΎΡΠΏΠ΅ΡΠΈΡΠΈΡΠ΅ΡΠΊΠΈΡ
Π°Π½ΡΠΈΡΠ΅Π» ΠΏΡΠΎΡΠΈΠ² npe-MSK ΠΌΡ ΠΏΠΎΠΏΡΡΠ°Π»ΠΈΡΡ ΠΈΠ΄Π΅Π½ΡΠΈΡΠΈΡΠΈΡΠΎΠ²Π°ΡΡ Π³ΠΎΠΌΠΎΠ»ΠΎΠ³ΠΈ MSK Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ
ΠΌΠ»Π΅ΠΊΠΎΠΏΠΈΡΠ°ΡΡΠΈΡ
ΠΌΠ΅ΡΠΎΠ΄Π°ΠΌΠΈ ELISA ΠΈ ΠΠ΅ΡΡΠ΅ΡΠ½-Π±Π»ΠΎΡΠΈΠ½Π³Π°. Π ΡΠΈΡΠΎΠΏΠ»Π°Π·ΠΌΠ°ΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΈ ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΡΡ
ΡΡΠ°ΠΊΡΠΈΡΡ
Π»ΠΈΠ·Π°ΡΠΎΠ² ΠΊΠ»Π΅ΡΠΎΠΊ ΠΌΠ»Π΅ΠΊΠΎΠΏΠΈΡΠ°ΡΡΠΈΡ
Π½Π°ΠΌ ΡΠ΄Π°Π»ΠΎΡΡ ΠΎΠ±Π½Π°ΡΡΠΆΠΈΡΡ Π±Π΅Π»ΠΊΠΈ, ΠΈΠΌΠ΅ΡΡΠΈΠ΅ ΠΈΠΌΠΌΡΠ½ΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΈΠΉ ΠΏΠ΅ΡΠ΅ΠΊΡΠ΅ΡΡ Ρ MSK Π ΡΠΎΠ²ΠΎΠΊΡΠΏΠ½ΠΎΡΡΠΈ Ρ ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΠ°ΠΌΠΈ ΠΏΠ΅ΡΠ΅ΠΊΡΠ΅ΡΡΠ½ΠΎΠ³ΠΎ Π°ΠΌΠΈΠ½ΠΎΠ°ΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΡΠΈ Π΄Π°Π½Π½ΡΠ΅ Π΄Π°ΡΡ ΠΎΡΠ½ΠΎΠ²Π°Π½ΠΈΠ΅ ΠΏΡΠ΅Π΄ΠΏΠΎΠ»ΠΎΠΆΠΈΡΡ Π½Π°Π»ΠΈΒΡΠΈΠ΅ ΠΎΠ±ΡΠΈΡ
Π°Π½ΡΠΈΠ³Π΅Π½Π½ΡΡ
Π΄Π΅ΡΠ΅ΡΠΌΠΈΠ½Π°Π½Ρ Ρ ΠΌΠΈΡΠΎΡ
ΠΎΠ½Π΄ΡΠΈΠ°Π»ΡΠ½ΡΡ
ΠΈ ΡΠΈΡΠΎΠΏΠ»Π°Π·ΠΌΠ°ΡΠΈΡΠ΅ΡΠΊΠΈΡ
Π»ΠΈΠ·ΠΈΠ»-ΡΠ ΠΠ ΡΠΈΠ½ΡΠ΅ΡΠ°Π· Π²ΡΡΡΠΈΡ
ΠΆΠΈΠ²ΠΎΒΡΠ½ΡΡ
Characteristics of the nuclear (18S, 5.8S, 28S and 5S) and mitochondrial (12S and 16S) rRNA genes of Apis mellifera (Insecta: Hymenoptera): structure, organization, and retrotransposable elements
As an accompanying manuscript to the release of the honey bee genome, we report the entire sequence of the nuclear (18S, 5.8S, 28S and 5S) and mitochondrial (12S and 16S) ribosomal RNA (rRNA)-encoding gene sequences (rDNA) and related internally and externally transcribed spacer regions of Apis mellifera (Insecta: Hymenoptera: Apocrita). Additionally, we predict secondary structures for the mature rRNA molecules based on comparative sequence analyses with other arthropod taxa and reference to recently published crystal structures of the ribosome. In general, the structures of honey bee rRNAs are in agreement with previously predicted rRNA models from other arthropods in core regions of the rRNA, with little additional expansion in non-conserved regions. Our multiple sequence alignments are made available on several public databases and provide a preliminary establishment of a global structural model of all rRNAs from the insects. Additionally, we provide conserved stretches of sequences flanking the rDNA cistrons that comprise the externally transcribed spacer regions (ETS) and part of the intergenic spacer region (IGS), including several repetitive motifs. Finally, we report the occurrence of retrotransposition in the nuclear large subunit rDNA, as R2 elements are present in the usual insertion points found in other arthropods. Interestingly, functional R1 elements usually present in the genomes of insects were not detected in the honey bee rRNA genes. The reverse transcriptase products of the R2 elements are deduced from their putative open reading frames and structurally aligned with those from another hymenopteran insect, the jewel wasp Nasonia (Pteromalidae). Stretches of conserved amino acids shared between Apis and Nasonia are illustrated and serve as potential sites for primer design, as target amplicons within these R2 elements may serve as novel phylogenetic markers for Hymenoptera. Given the impending completion of the sequencing of the Nasonia genome, we expect our report eventually to shed light on the evolution of the hymenopteran genome within higher insects, particularly regarding the relative maintenance of conserved rDNA genes, related variable spacer regions and retrotransposable elements