WASHINGTON UNIVERSITY TU€ HENRY SHAW SCHOOL OF BOTANY SAINT LOUIS March 15, 1955 Dr, Rosalind E, Franklin Birkbeck College University of London 21 Torrington Square, W,C.l, London England Dear Dr, Franklin: deal of interest, a great deal of what we want to know about the relations between TMV and the nonvirus proteins. I have gone over the material which you send me with a great Your investigations certainly promise to tell us Although there are a number of extremely interesting points which I shall want to write you about, this letter will be restricted to the immediate issue of the paper which you have prepared on your results with B8. After reading your manuscript it appears evident that a good deal of confusion now exists with regard to the abnormal proteins related to TMV. The facts as I see them are these, 1) W-infected plants produce at least two and'probably three low-molecular weight abnormal nonvirus proteins which are immuno- chemically related to TMV and which polymerize to form "V-like rods. As noted in my revision of your manuscript, there is agreement on the existence of at least two of the above proteins between myself, Jeener and Takahashits ori inal publication (her, Jour. of Botany, 40, 85) , %he correlations + etween my terminology and Takahashits, with respect to the two proteins, is presented in my revision of your manuscript, 2) 3) Very recently the paper by Delwiche, Takahashi, Newmark, et al, (Biochim. Biophys. Acta l6, 127) claims that only a single abnormar - protein can be found, However, they offer no data to substantiate this claim and the method of preparation which they use is identical with one of the methods which we have employed to isolate two separate non- virus oomponents (B3 and B6), been concerned over this discrepancy and have examined the behavior of mixtures of B3 and B6. a single high-molecular weight component which migrates in a homogeneous fashion in the Pbelius apparatus at pH 7.0, components will form a co-polymer, made by first preparing a homogeneous preparation of B3 and then polymerizing it to form B8, polymerization is induced it is likely that a complex co-polymer will be formed which is difficult to distinguish from B8. Since seeing the Delwiche paper I have We find that such mixtures polymerize to form In other words, the two Our own preparations of B8 are Unless B3 and B6 are separated before I suspect very -2- March 15, 1955 strongly that the protein described by Delwiche et al, and which was used by Rich in his work vas very likely such a =-polymer containing both B3 and B6 in an indefinite ratio, 4) For these reasons the Babnormal proteinn of Rich and Delwiche should not be considered the equivalent of B8. workers, including Takahashi, are not aware of the occurrence of B6 and its participation in the formation of a co-polymer, it is entirely likely that successive preparations from their laboratories may contain variable ratios of B3 and B6, equivalent to our B8, extracts are manipulated and it is possible to lose most of the B6 in a preparation as a result of lengthy processing, that the major variation in the preparations produced by the other laboratories would be a fluctuation in the relative amount of B6 present in the polymerized "abnormal proteinw and that in some cases the B6 content may be zero without their being aware of it, your manuscript, that you will feel free to condense it as far as possible, also made a few minor changes in the first page or two of your manuscript to avoid statements which cannot be supported by our own data, This includes Richst assertion that the chemical composition of the abnormal protein and TMV are identical, statement in view of the likelihood that their protein is not of constant composition, Furthermore, if these Thus, the reparation which Takahashi has sent you may contain little or no B z and is, therefore, We find that B6 is readily denatured when Hence, I would expect 1 have tried to incorporate some of these ideas in an addition to What I have added is probably unduly long and I hope I have This is a very doubtful Because of the material which I have added to the manuscript I think that it would be wise if my name were added to the paper. !€'he results are extremely interesting and I am happy for the opportunity to be associated with the publication, Also included herewith are galley proofs of two papers on the nonvirus proteins which will appear in the March 20th issue of the JOURNAL OF GENERAL PHYSIOLOGY. can have idea of the evidence which we now have on the wU%&&?- rhd and properties of the nonvirus proteins, I am sending you these now so that you some Under separate cover I am sending you about 10 milligrams more of the B8 preparation which you now have, After some trials I decided that the best orientated preparation appeared to be a pellet of the protein spun down in the ultracentrifuge. The material is being sent to you in this form, in particle alignment by forcing this material into a capillary, any event, the pellet which I am sending you is the most concentrated form in which I can prepare B8 and should make your manipulations easier to carry out. I hope that you will be able to get an improvement In -+d-L- -3 - March 15, 1955 We are very much interested in obtaining good amino acid analyses on this preparation of B8, try to salvage as much material as you can from your operations and return it to me when it is no longer useful to you. Regardless of the physical state of the preparation we can use it for amino acid anal ysi 8 , Consequently, I should like to ask you to I hope you will write again soon and let me know what you think of the situation, In a little while I shall write you concerning some of the ideas which I have on the possible relations between the abnormal polymerized proteins and TMV nucleoprotein, Meanwhile, 1 shall look forward with interest to learning of your latest results. Sincere yours, 3-cw. Barry Commoner BC: bl Encl ,