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• W115881036 abstract "The steady-state levels and metabolic properties of mitochondrial tRNAshave been analyzed in Hela cells and correlated with the function of the tRNAsfor organelle-specific protein synthesis. DNA excess hybridization experimentsutilizing separated strands of mitochondrial DNA (mtDNA) and purified tRNAsamples from exponential cells long-term labeled with [^(32)P] orthophosphate haverevealed a steady-state level of 6 x 10^5 tRNA molecules per cell, with threefourthsbeing encoded in the heavy (H)-strand and one-fourth in the light(L)-strand. Hybridization of the tRNAs with a panel of M13 clones of humanmtDNA containing, in most cases, single tRNA genes and a quantitation of two dimensionalelectrophoretic fractionations of the tRNAs have shown that thesteady-state levels of tRNA^F and tRNA^V are two to three times higher than theaverage level of the other H-strand-encoded tRNAs and three to four times higherthan the average level of the L-strand-encoded tRNAs. Similar experimentscarried out with tRNAs from cells labeled with very short pulses of [5-^3H] uridinehave indicated that the rates of formation of the individual tRNA species areproportional to their steady state amounts. Therefore, the 15-fold to 60-foldhigher rate of transcription of the tRNA^V and tRNA^F genes (transcribed with therDNA transcription unit) relative to the other H-strand tRNA genes (transcribedwith the whole H-strand transcription unit) and the 13-fold to 20-fold higher rateof transcription of the L-strand tRNA genes relative to the H-strand tRNA genes(other than tRNA^V and tRNA^F genes) are not reflected in the rates of formationof the corresponding tRNAs. The available data indicate that the majority oftRNA^V and tRNA^F transcribed from the rDNA transcription unit are degraded asthey are excised from the primary transcripts. It also seems likely that themajority of the L-strand-encoded tRNAs are degraded before they are excisedfrom the short-lived polycistronic transcripts. Furthermore, a role of the aminoacyl tRNA synthetases in stabilizing the different tRNA species atrelatively uniform levels is suggested. A comparison of the steady-state levels ofthe individual tRNAs with the corresponding codon usage for protein synthesis, asdetermined from the DNA sequence and the rates of synthesis of the variouspolypeptides, has not revealed any significant correlation between the twoparameters. In other experiments, isolated human mitochondria containing amitochondrial DNA (mtDNA)-coded chloramphenicol resistance marker wereinjected at an average dose of less than one into sensitive human cells partiallydepleted of their mtDNA by ethidium bromide treatment. Under selectiveconditions, the mitochondria became established in the recipient cells with afrequency greater than 2 to 3 x 10^(-3). A rapid and, in some cases, completereplacement of the resident mtDNA by the exogenous mtDNA took place in thetransformants, as shown by multiple mtDNA and nuclear DNA polymorphisms.Intracellular mtDNA selection played a crucial role in this replacement, withsignificant implications for mitochondrial genetics." @default.
• W115881036 created "2016-06-24" @default.
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• W115881036 date "1988-01-01" @default.
• W115881036 modified "2023-09-23" @default.
• W115881036 title "Studies of human mitochondria. Steady-state levels and metabolic properties of the mitochondrial tRNAs. Injection of mitochondria into human cells leads to a rapid replacement of the endogenous mitochondrial DNA" @default.
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