NEWS

Oscar Miller (1925 – 2011) – the technological genius of chromosome science


Oscar and his wife Mary-Rose 1973

Oscar Miller died on January 28, 2012 at the University of Virginia Hospital.
He was born on April 12, 1925 in Gastonia, North Carolina. He served in the US Navy from 1943 through 1946. After earning a bachelor's and master's degrees in agronomy from North Carolina State University, he was a farmer for six years and then enrolled in the University of Minnesota where he was awarded a PhD in plant genetics. He joined the research staff at Oak Ridge National Laboratory in 1961 and began the work that earned him worldwide renown as a molecular biologist.
Oscar was elected to the National Academy of Sciences in 1978. He was named a Fellow of the American Association for the Advancement of Science in 1980. He was Chairman of the Department of Biology and held the William R. Kenan and the Lewis and Clark Professorship at the University of Virginia. He was visiting professor of biology at the Center of Investigation and Advanced Studies in Mexico City, the California Institute of Technology, the Max-Planck-Institut für Zellbiologie in Heidelberg, Germany and the University of California at Irvine. He was the Senior Fulbright Scholar at the Division of Molecular Biology, CSIRO at New South Wales, Australia. Among his many honours from around the world, he received the Life Achievement Award in Science from the Commonwealth of Virginia in 1997.
Prior to Oscar Miller's time, the existence of genes and of the double helical structure of DNA were accepted in the scientific community as sound hypotheses but what genes did and how they did it in terms of transcription was beyond the scope of actual observation. Then Oscar developed a technique – now known as "Miller spreading"- that enabled scientists, through electron microscopy, to visualize individual genes caught in the very act of making their RNA transcripts and this, of course, opened the door to many important advances in chromosome and nuclear science.
I first met Oscar in 1969, shortly after he had developed and published his spreading technique. I remember being utterly humbled and overawed by the extraordinary skill, ingenuity, inventiveness and courage behind the work. His method is described in his 1969 Science paper (with Barbara Beatty) as follows:

The specimen was prepared by placing the contents of an oocyte nucleus in deionised water, thus causing dispersal of the nucleolar components; the unwound cores were centrifuged through a neutral solution of 0.1 molar sucrose with 10% formalin onto a carbon covered grid; the grid was rinsed in 0.4% Kodak Photo-Flo solution before drying; the preparation was then stained for 1 minute with 1% phosphotungstic acid in 50% ethanol at pH 2.5 (unadjusted), rinsed in 95% and then 100% ethanol, and dried with isopentane. (Miller, O.L. and Beatty B.R. 1969. Visualization of nucleolar genes. Science N.Y. 164, 955 – 957)

An electron micrograph of a single nucleolar gene transcription unit prepared by the Miller spreading technique. c: non-transcribing chromatin of the spacer region. p: the position of the first polymerase molecules at the start of the transcription unit. t: closely packed RNA transcripts extending sideways from the DNA axis. e: the end of the transcription unit.

Modern young chromosome scientists can have no concept of the reality of the process outlined in this brief description. In culinary terms, it was like making the world's most delicious cake "from absolute scratch". First, isolate the oocyte nucleus - not the easiest thing to do. Isolate it in deionised water – well, what you don't know is that it immediately disappears, so what follows is done on faith. Centrifuge the contents through a sucrose/formalin mix – lots of ingenuity and testing to find the right concentrations. Centrifuge the contents onto an electron microscope grid. Well, the grid is 3mm in diameter and extremely delicate and the whole business depends on scrupulous attention to detail and cleanliness – and again, lots of trial and error. Carbon covered grid – you could spend days learning to make really good carbon coated grids! Besides, centrifuge - with what? You couldn't buy a centrifuge for spinning DNA onto grids – you had to make one. And when you had done that you had find out how fast to spin and how long to spin and you had to find out the hard way that if you started the spin too fast or too slow, everything landed up in a heap at one edge of the grid. We tried to extend Oscar's work to look at transcription on the loops of the lampbrush chromosomes in amphibian oocytes and we discovered just how much skill and patience is needed to produce meaningful results – and we had the benefit of tapping into Oscar's experience. I remember well my first reaction when I saw the pictures of his transcribing genes and saw how he had done it: this man was truly a technological genius. Since its original publication in 1969 the Miller and Beatty paper has been cited nearly 1000 times. In terms of "impact" in the field of chromosome and nuclear science it is almost unrivalled.
The Oscar I remember was a great character - in that smooth, quiet, Southern and utterly charming way, all on top of a supreme competence as a scientist and teacher. Those of us who knew him will feel a sense of privilege and pleasure. Young chromosomologists the world over should take time to look at some of his work and perhaps aspire to be just a little bit like him. Science "from scratch" is fun and Oscar was one of the best at doing this kind of thing.

Herbert Macgregor

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January 2008

A new study of transcription on lampbrush chromosomes from the Morgan Laboratory in Nottingham, England. Just published in Chromosome Research vol. 15, pages 985 - 1000 .

Localized co-transcriptional recruitment of the multifunctional RNA-binding protein CELF1 by lampbrush chromosome transcription units. Chromosome Research volume 15 pages 985 - 1000.

A skilful explotation of the highly extended transcription units of LBCs to investigate whether CELF1, an RNA-binding protein involved in the regulation of alternative splicing, mRNA stability and translation, is localized to active transcription units. Multiple functions of CELF1 explored in this remarkable system and some nice new ideas forthcoming.

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January 2008

From Michel Bellini December 20th 2007 - TO BE PUBLISHED IN CHROMOSOME RESEARCH VOLUME 16 NUMBER 2

Over the last two years we have been optimizing a method to observe
lampbrush chromosomes in oil-isolated GVs from stage IV-VI oocytes,
which I think is as close as it gets to an in vivo situation.  We
obtained beautiful images of live LBCs by phase contrast (or DIC) and
DAPI staining.  Interestingly, while the axes of both homologous
chromosomes of an LBC are readily observable, we yet have to see the
lateral loops.  We were also able to demonstrate the recruitment of
several chromosomal proteins tagged with YFP.  One of them, MCD1 (a
member of the cohesin complex) associates with the chromosomal axes
and we are analyzing now its kinetic behavior by FRAP. 

We are now trying to solve why we cannot see the chromosome loops in
oil-isolated GVs.  We are currently tagging several loop proteins
with GFP (and soon Photo-activable GFP) to try to visualize for the
first time and at a high spatial resolution a PolII transcriptional
unit live.

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