The Mitochondrial Free Radical Theory of Aging - Supernova: Pliki

The Mitochondrial Free Radical Theory of Aging - Supernova: Pliki The Mitochondrial Free Radical Theory of Aging - Supernova: Pliki

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An Introduction to Mitochondria Fig. 2.8. The chemiosmotic proton circuit. laboratory at a particular time. The discovery was that mitochondria contained their own DNA, and the best that can be said with fairness is that it spanned a period between 1959, when Chèvremont and colleagues first reported evidence for it, 30 and 1963, when Nass and Nass published a clear demonstration that a frequently isolated component of mitochondria was indeed DNA. 31 No one had any reason to suppose that mitochondria needed DNA, so there was initially a tendency to ascribe its presence in mitochondrial preparations to nuclear contamination. Indeed, mitochondria are the only components of animal cells’ cytoplasm that have any of their own DNA;* all the rest of our genetic material is stored in our chromosomes, in the nucleus. This reticence was somewhat compensated in the following years. There was still a lingering suspicion that the mitochondrial DNA might be of nuclear origin after all, but this was swept away by the discovery that it is a circular molecule, in contrast to the linear character of all eukaryotic chromosomes. This discovery was made in 1966, and published with alacrity, by three labs independently. 32-34 Each mitochondrion has several copies of the mtDNA. The exact number varies between species; in humans it is typically about five. 2.4.2. Inheritance of mtDNA Erythrocytes are the only mammalian cells with no mitochondria, but there is another cell type which has very few: sperm. Mitochondria are passed on from one generation to the next in egg cells, but almost none are contributed by sperm.** This is largely because sperm * There is still some debate on this point with regard to centrioles, but the view that they have their own DNA is not currently favoured. ** It is widely stated that no mtDNA whatever is paternally inherited, but this is incorrect: see Ref. 35 for an eloquent dissection of this “meme.” 21

22 The Mitochondrial Free Radical Theory of Aging are so small, containing under 100 mitochondria (located in the tail); also, there may be a system whereby any paternal mtDNA that gets into the fertilized egg is then actively destroyed. 36 This situation is of course in complete contrast to the way that the rest of our DNA is inherited, namely that we receive one copy of each chromosome from each parent (with the exception of the sex chromosomes). The almost solely maternal inheritance of mitochondrial DNA has proved very useful in many areas of science: it has helped us to discover the patterns of migration of early man, 37 and also to pin down the genetic cause of certain degenerative diseases, which were found to be maternally inherited and then to be associated with particular mitochondrial DNA mutations (see Section 6.6.5). 38,39 2.4.3. The Nuclear-Coded Parts of Mitochondria; Protein Import Quite soon after mtDNA and its maternal inheritance were discovered, genetic studies began to show that some of the enzymes known to be present in mitochondria exhibited normal Mendelian inheritance, which meant that their genes must be carried in the nucleus, not in the mtDNA. In 1981, when the human mtDNA was sequenced, 40 it was established that just 13 proteins are encoded in it—a small fraction of those known to be mitochondrially located. The hundreds of other proteins that are found in mitochondria are all encoded in nuclear, chromosomal DNA, and are constructed in the cytoplasm just like all other chromosomally encoded proteins. They are then imported into mitochondria. The import mechanism is highly sophisticated, and is still the subject of intense study. It carries molecules across the energy-transducing inner membrane, so it must be as fastidious as all the other carrier systems described earlier (see Section 2.3.2.4) in not allowing free transit (“leakage”) of protons or other ions; but it must do so while translocating a molecule far larger than is carried by those other systems. Also, since proteins are too big to fit through the pores of the outer membrane, they must be dragged through both membranes. This has traditionally been supposed to occur at specialised membrane locations, the “contact sites” mentioned earlier, but there is now good evidence that the protein translocation systems of the outer and inner membranes are separate except when engaged in translocation: 41 the contact sites are indeed the sites of import, but they are created by the import process pulling the membranes together. (This is why they were not shown in Figure 2.2.) The proteins involved are all found to be located strictly in one or the other membrane (or in the intermembrane space), and are in fact now named accordingly—“Tom” and “Tim” (followed by a number indicating the particular protein’s molecular weight) for “Transporter of the outer (inner) mitochondrial membrane.”* The channel formed by the Tom proteins is also often termed the general import pore. One of the first things to be discovered about the import process was the characteristic by which a protein destined for the mitochondrion is recognised as such. Nearly all of those initially studied have a “signal” sequence of a few dozen amino acids, at the beginning of the protein, which has no role in the protein’s ultimate function—in fact, it is chopped off once the protein is safely inside the mitochondrion. 42 But prior to import, it acts to tell the cell’s transport machinery to take it to a mitochondrion. Experiments have shown that typical cytosolic proteins (ones that are not normally imported into mitochondria) will be imported into mitochondria if such a signal is attached to them. 43 * This nomenclature, due to Nikolaus Pfanner, is so attractive that it has been copied by botanists to describe the corresponding proteins of chloroplasts, which are called Tocs and Tics. Some primitive organisms have organelles with three or four membranes; I await the corresponding nomenclature with interest.

An Introduction to Mitochondria<br />

Fig. 2.8. <strong>The</strong> chemiosmotic proton circuit.<br />

laboratory at a particular time. <strong>The</strong> discovery was that mitochondria contained their own<br />

DNA, and the best that can be said with fairness is that it spanned a period between 1959,<br />

when Chèvremont and colleagues first reported evidence for it, 30 and 1963, when Nass and<br />

Nass published a clear demonstration that a frequently isolated component <strong>of</strong> mitochondria<br />

was indeed DNA. 31 No one had any reason to suppose that mitochondria needed DNA, so<br />

there was initially a tendency to ascribe its presence in mitochondrial preparations to nuclear<br />

contamination. Indeed, mitochondria are the only components <strong>of</strong> animal cells’ cytoplasm<br />

that have any <strong>of</strong> their own DNA;* all the rest <strong>of</strong> our genetic material is stored in our<br />

chromosomes, in the nucleus.<br />

This reticence was somewhat compensated in the following years. <strong>The</strong>re was still a<br />

lingering suspicion that the mitochondrial DNA might be <strong>of</strong> nuclear origin after all, but this<br />

was swept away by the discovery that it is a circular molecule, in contrast to the linear character<br />

<strong>of</strong> all eukaryotic chromosomes. This discovery was made in 1966, and published with alacrity,<br />

by three labs independently. 32-34<br />

Each mitochondrion has several copies <strong>of</strong> the mtDNA. <strong>The</strong> exact number varies between<br />

species; in humans it is typically about five.<br />

2.4.2. Inheritance <strong>of</strong> mtDNA<br />

Erythrocytes are the only mammalian cells with no mitochondria, but there is another<br />

cell type which has very few: sperm. Mitochondria are passed on from one generation to the<br />

next in egg cells, but almost none are contributed by sperm.** This is largely because sperm<br />

* <strong>The</strong>re is still some debate on this point with regard to centrioles, but the view that they have their own DNA<br />

is not currently favoured.<br />

** It is widely stated that no mtDNA whatever is paternally inherited, but this is incorrect: see Ref. 35 for<br />

an eloquent dissection <strong>of</strong> this “meme.”<br />

21

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