Subnuclear components. preparation and fractionation

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TIBS - February 1977 I

Subnuclear Components. and Fractionation

BOOKS Marginally review

useful

Genetics and Biogenesis of Mitochondria and Chloroplasts edited by C. William Birky, Jr, Philip S. Perlman and Thomas J. Byers, published by Ohio State University Press, Columbus, Ohio, 1976. $IS.- (approx. .E8.-) (x-1-361 pages)

The book is a collection of ten papers submitted by the lecturers attending a colloquium held in 1974 at Ohio State University. In the preface to the book J. P. Kreier and R. 0. Moore state the aim of the colloquium was ‘to provide a forum in which research workers from the college and other institutions would present their work to each other and to interested third parties in a direct and personal way’. This is certainly an intelligent and noble target and I am sure that the meeting succeeded in fulfilling the expectations of the organizers. What I personally object to is the usefulness of publishing what appear to be greatly enlarged or reduced versions of the lectures presented by the different speakers. Indeed, it seems to me that the published contributions differ widely in length and depth. For instance, Sager’s paper, dealing with the possible existence of a restriction-modification mechanism in uniparental inheritance, seems to be an abridged version of a paper published last year in Science. Conversely, Attardi presents an exhaustive (and exhausting, 63 pages out of a total of 352) treatment of the biogenesis of mitochondria in HeLa cells. The paper is limited to the presentation and the discussion of Attardi’s results over the past few years. This is quite alright as his group is one of the leading teams studying mitochondrial biogenesis and possibly the spearhead in the case of mitochondria from cultured mammalian cells. My second complaint is that to read something novel one has to sift through tens of pages of published data already discussed in other reviews, contributions to symposia, etc. The same considerations may be applied to Mahler’s paper, another 50-page hunk. Here again,, one of the leading scientists in the molecblar biology of

Nuclear fractionation a critical look

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Preparation

by G. D. Birnie, published by Butterworths, London, 1976. &15.- (approx. $28.-) (334 pages) yeast mitochondria, re-elaborates dat;a that he himself has already discussed in a.t least six reviews or books in the last threje years. Yeast mitochondria are also thle subject of Grifflths’ paper, dealing with th e use of antibiotic-resistant mutants to disIsect the pathway of oxidative phosphor .ylation, and the contribution of Perlmal n on cytoplasmic petite mutants. The genetics of mitochondria in fung and ciliates is treated by Birky whil Laughnan and Gabay deal with cytoplas mic pollen sterility in maize, presenting th argument that the phenomenon may b caused by an episome-like factor. Besides the contribution by Sager, th.e ‘chloroplast side’ is covered by Hoobe :r and Stegeman with a paper on the charac :teristics and the site of synthesis of th:e thylakoid membranes in Chlamydomonfi IS reinhardii, by Tilney-Bassett on th.e genetics of variegation in higher plantl $3 and by Wildman summing up his work o n the origin and evolution of two chloroplar ;t proteins, namely ferredoxin and Fractio n I protein. The latter paper is, in my op inion, fascinating reading for the molecula n biologist as well as for the student of plar It taxonomy or evolution. As in many books collecting paper ‘S from different authors, the index appear ‘S to be inadequate and, at times, misleadinl 3. Having a personal interest in ribosome! % I did look for a relevant !leading. I foun d only a cross-reference in ‘Protein syr Lthesis, polysomes. See Ribosomes, mitt )chondrial’. However, ribosomes do nc,t appear in the index and, under ‘Mitochor Idria’, one sees ‘See specific topics’. Sim llarly, one finds ‘Fraction I protein. St se Ribulose diphosphate carboxylase’ bu t, alas, this enzyme is not mentioned in thLe index. In the last few years, reviewers have bee n complaining, with reason, about the rocl ceting prices of scientific books. Pit US$lS.-, this book does not appear to t be too expensive and it may be a good bu‘Y for the library not too well stocked in thle above-mentioned fields. OR10 CIFERI U 0. Ciferri is Professor of Microbiology, University Of Pavia, Pavia, Italy.

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This is the natural and long expected sequel of the successful ‘Subcellular Components : Preparation and Fractionation’ by the same author, which has served and still serves so many students and researchers using cell fractionation techniques both as a guide and a ‘cookbook’. The present book focuses on the isolation and characterization of nuclear structures and molecule classes, and most of the articles present an unusually critical evaluation of the methods that are currently used in the particular field. The emphasis of the whole book is on critical comparison of the various procedures and on the value of the criteria used to assess the purity and the structural integrity of fractions. Thus it invites the reader to think somewhat dialectically and to weigh up the pros and cons of a specific method rather than simply to follow the author’s favourite techniques and concepts. In general, the literature is adequately evaluated up to 1974. One of the highlights of the book is certainly the very carefully written article by D. Rickwood and G.D. Birnie on the various ways to isolate, fractionate and characterize chromatin, which has to be seen in context with two subsequent, similarly detailed chapters on histones (E. W. Johns) and non-histone proteins (A. J. McGillivray). These authors are also not reluctant to touch some of the ‘hot themes’ and, for example, seldom has the questionable validity of so-called transcriptionally ‘active’ and ‘inactive’ chromatin subfractions so clearly been examined as in this contribution by Rickwood and Birnie. Two articles deal with methods for extracting RNA (M.E. Bramwell) and DNA (P.H. Butterworth) from whole cells, nuclei and nuclear subfractionations. While the biochemical methods used to characterize the size and purity of such preparations are well compiled, the value of the electron microscopic methods in this field seems to be underrated by the authors. U. E. Loening and A.M. Baker have contributed a sovereignly written review on the isolation of nucleoli from animal and plant cells and have focused on the recent developments in this field, (i.e. work later than the classic review in the ‘Nucleolus-book’ by Busch and Smetana). An unusually critical chapter is that by D.J. Fry on isolated nuclear envelope

TIBS - February 1977

46 material. This author has compared, with scrupulous exactitude, nearly all procedures hitherto published in the literature and offers a series of very valuable conclusions and advice on which method should be used for which specific question in the subsequent analysis of the nuclear membrane fractions. The introductory chapter by E.A. Smuckler, M. Koplitz and D. E. Smuckler gives a brief summary of nuclear morphology and presents the various principles in methods of isolating animal nuclei (plant material is indeed largely underrepresented). However, this article in particular, but to some degree the whole book, suffers from the unsatisfactory print quality of the electron micrographs. Apart from this deficiency, the book is such a valuable compilation and critical guide that one can hardly imagine a biologist or biochemist working with nuclei who can afford not to have it. WERNER W. FRANKE W. W. Franke is head of the Division qf Membrane Biology and Biochemistry at the Institute of Experimental Pathology, German Cancer Research Center, Heidelberg, G. F. R.

Flavins and Flavoproteins edited by T. P. Singer, published by Elsevier, Amsterdam, 1976. Djl.258.($99.50) (xiii+813pages)

In the spring of 1975 the Fifth International Symposium on Flavins and Flavoproteins was held in San Francisco. The original meaning of a symposium was an Ancient Greek drinking party, and although in this instance there were no oldAthenian participants, the editor makes it clear in his introduction that a very good time was had by all. In fact, like many ‘international’ meetings, the majority of contributions came from the U.S.A., but with respectable offerings from Europe and Japan. During four days, 92 original papers were presented and these, apart from three too late for inclusion, have each a ‘chapter’ in this large volume, grouped conveniently into thirteen parts. The editor and host, T.P. Singer, provides a useful introduction, incidentally revealing the familiar but often unadmitted truth that enthusiasm for a chosen research area derives as much from aesthetic satisfaction as fr,jrn the conventional scientific motivations.

Now for the papers. We start in at the deep end with oxygen activation, papers which are hard going except for specialists, but with excellent presentations and discussions. The following ‘Bacterial Luciferase’ papers are more congenial, directed mainly at elucidating the reaction intermediates, and all agree that FMN is not itself the bioluminescent emitter. Good summaries make rapid comprehension easier and here, and throughout the book, one is impressed by the capability and skilful use of the many physical techniques employed. Interest is evenly divided between the macromolecular properties and reaction pathways of oxygenases (mostly hydroxylases) in the following eight papers, impressive in the wide variety of reactions catalysed by these enzymes. Next, we explore proton abstraction processes in several well-defined, enzyme-substrate systems, mostly down-to-earth stuff, except for one paper devoted to bio-organit’ mechanisms, and another on chargetransfer complexes. Covalently-bound ‘old’ and ‘new’ flavins are investigated in a larger group of papers, followed by an intriguing selection of studies using flavin analogues in which N is replaced by C or S. The later papers in this section have formidable but informativeltreatmentsof the intimate electronic structure of the flavin nucleus. The molecular structure and binding properties, redox mechanisms and molecular evolution of flavodoxins comprise the following section. That old favourite, lipoyl dehydrogenase, has a central position in this book, as it has in the hearts of the older flavinophiles. Active site properties and reaction mechanisms are of prime interest here, and obviously this old friend still has much to reveal. We then come to complex metallo-flavoproteins, in papers much more of the ‘enzyme isolation and characterization’ type previously rare in this8volume.Adrenodoxinsand flavoprotein oxidases have each a small group of contributions, and, nearing the end of the book, we swing away from the biophysical and mechanistic approach hitherto dominant, and conclude with riboflavin biosynthesis and metabolism; and a more heterogeneous group of unclubbable ‘special topics’. Taken altogether, the standard of the papers is remarkably high with many impressive examples of the power of the newer biophysical, especially spectroscopic, techniques in the flavin field. In many cases they would serve as excellent models for research workers who are unfamiliar with these experimental approaches. Although it is regrettable that summaries are not always provided, the discussions are almost always stimulating and

should reassure those who may doubt that we have much more to learn about flavoproteins that innumerable questions remain as yet unanswered. The text is well illustrated and remarkably free from printing errors (I counted about twenty in the whole book), and the index is reasonably helpful. As with all such collections, some of the material has been published elsewhere, but appearing as it does relatively soon after the symposium, this is not a serious shortcoming. Despite the formidable cost of this book, the quality and breadth of the material gathered together in this collection make it an essential acquisition by any biochemical or biophysical laboratory actively engaged or interested in the flavin field. F. W. J. TEALE F. W. J. Teale is the Reader in Physical Biochemistry at the University #Birmingham, Edgbaston. U.K.

Skin deep The Life That Lives on Man by Michael L. A. Andrews, published by Faber & Faber, London, 1976. &4.95 (approx. $9.-) (183 pages)

There are moderate-sized tree trunks every thirty meters. They arise from greasy pits. Scattered over the surface are irregular salt-encrusted rims of drain-pipes disgorging droplets of water several meters across at the slightest rise in temperature. The inhabitants of this savanna are square in appearance and about the size of a shrew. They stretch and make a crackling sound and break into two individuals every half an hour or so. If you could look at your skin at an enlargement of 10000 times, this is how you would see your hairs, your openings of sweat glands and your bacteria. The healthy human skin is an ecosystem, just like a pond or a wood. In an environment of keratin soil with hairs and sweatgland discharge, a dense bacterial flora not only breaks down different components (causing, among other things, nasty smells) but also protects us against pathogenic species. On less healthy skins, we could find fleas, head- or crab lice drinking their blood meals, and sometimes even an itch mite taking great pains in burrowing just under the keratin layer. Other arthropods are found on healthy skins but are vectors of pathogenic organisms. On less populated or damaged skin the bacterium Staphylococcus aureus causes much destruction. Sometimes warts arise as a host action governed by a Papova virus. Much more happens on our skin, healthy or sick, and Andrews tells us about