From owner-biophysics@net.bio.net Sun Sep 13 23:00:00 1998
Path: biosci!biosci!not-for-mail
From: "clive delmonte" <clived@ndirect.co.uk>
Newsgroups: bionet.biophysics,bionet.molec-model,bionet.xtallography
Subject: DNA Structure Discussion Continued
Date: 14 Sep 1998 10:56:30 -0700
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Xref: biosci bionet.biophysics:4396 bionet.molec-model:2249 bionet.xtallography:4399

Bill further asks whether the side-by-side structure set out in my first
attachment does not contradict the structures derived from the fibres.  This
is a key question and requires a full answer (if you'll indulge me a
little.)  There are several aspects to this.  First, Crick had never seen
any of Franklin's (high quality) DNA fibre diffraction patterns when he and
Watson proposed their structure.  Watson had seen a B-DNA pattern only very
briefly and could not study it at leisure.  Plectonaemic winding in DNA had
already been proposed by Pauling & Corey early in 1953 and Watson & Crick
seem to have adopted that idea.

A R Stokes, a colleague and co-worker of Crick, had deduced an important
geometric ratio for the classical helical cross (Prog. Biophys. & Biophys.
Chem. 5 (1955) 140-167, page 163) (see attachment) such that, for the angle
from the vertical (delta), tan delta = helical pitch / (helical
circumference).  In Franklin's famous B-DNA diffraction pattern (see
attachment), delta = 40 degrees approximately, and, as the pitch is known to
be 33.4 Angstroms from the layer lines, we deduce that the major diffractors
lie on a helix of diameter of about 12 to 13 Angstroms.  This is close to
Franklin's deduction of a diameter of 11 Angstroms from her Pattersons.

I now have seventeen published DNA fibre diffraction studies in which the
researchers make it explicitly clear that they chose a unit cell which would
be large enough to accommodate a double helical diameter of some 19-23
Angstroms even though their patterns of systematic absences in the
reflections would have allowed them to choose a smaller unit cell (into
which the double helix could not be fitted.)  I also have references to
unpublished studies in 3 or 4 PhD theses where a larger unit cell was chosen
than the raw diffraction data actually required.  I should be very glad to
forward the list of references to anyone interested.

If you choose the smaller unit cell allowed by the dffraction pattern and
recalculate the helical diameter (a simple geometric exercise, of course),
you get diameters in the range 11.2 to 13.6 Angstroms.

A really fascinating study in conventional chemistry is that of James &
Mazia (Biochim.Biophys.Acta 10 (1953) 367-370).  They discovered that 1 mg
of dry calf thymus DNA could be spread in a monolayer to cover 0.28 square
metres.  They didn't notice that from just this result alone we can directly
estimate the minor axis diameter of the duplex as 11.7 Angstroms.  I would
be very glad if people would like to calculate the diameter for themselves
and compare their sums with mine.  (James & Mazia measured the major axis
diameter, that is the height of the spread film in a separate experiment as
21.6 Angstroms.  Therefore their results lead us to a duplex with an
elliptical section.)

The new model for DNA structure leads to the presence of a "front" and a
"rear" face with great implications for protein-DNA recognition, including
the way such DNA would wind onto nucleosomal cores, for example.

What problems would people see with this account so far, please ?
Perhaps from NMR or diffraction from true crystals of oligonucleotides ?
clive delmonte
clived@ndirect.co.uk




begin 666 Pagis Document (4).xif
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end


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