Research in Microbiology 170 (2019) 117e120
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Research in Microbiology
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In Memoriam
Obituary: Agnes Ullmann (1927e2019)*
« Where are you from? » « Transylvania, she would answer ». A
country that does not exist! Just a region, now part of Romania, but
which belonged intermittently to Hungary, Romany, and Germany
and was even part of the Ottoman Empire. She spoke French and En-
glish, but with a slight accent that betrayed her origin. She also spoke
Hungarian, her mother tongue, as well as Romanian and German.
Agnes Ullmann was born in 1927. She was twelve when World
War II began. At that time, she spent much of her time on sport
fields. From then on, she continued practicing various sports
including swimming and, when she could, mixed science and her
passion for sports. Thus, she managed to participate for years in
the “Rencontres de Me
ribel” a one-week workshop in the French
Alps, combining early morning and late afternoon lectures with
ski from 11 am to 5 pm. When she became unable to practice sports,
she followed some of them with passion. During her last years, it
was not proper to call her on the phone when she was watching
a tennis competition such as Roland Garros on the television!
After the war, she attended the Faculty of science at the Univer-
sity of Cluj, a town then in Hungary but which became Romanian
after the Paris peace treaty in 1947. Even though the postwar period
was not easy and she was often hungry and cold, she was then
immersed in a climate of culture, including music, for which she
developed a profound love that never left her. However, by that
time, she had decided that she wanted to become a scientist, and
science instruction was far from satisfactory at that university.
Therefore, she decided to continue her formation at the University
of Budapest, leaving Romania for Hungary. There, she obtained her
*
This article is greatly inspired from an article published by Agnes Ullmann enti-
tled “A Fortunate Journey on Uneven Grounds” (Annu. Rev. Microbiol. 2012. 66:
1e24). This article includes references to her work. Vivid accounts from the life
in Hungary when Agnes was living there, and from her escape from this country,
can be found in the book Brave Genius, published in 2013 by Sean B. Carroll (Crown
publishers, New York).
https://doi.org/10.1016/j.resmic.2019.04.001
0923-2508
chemistry diploma in 1949. Then she started to work in the Depart-
ment of Medical Chemistry, directed by Bruno F. Straub, a former
student of the Nobel Prize winner Albert Szent-Gyo €rgyi. Everyday
life was very difficult. Hungary then experienced the hardest dicta-
torship in Europe. Agnes was among the thousands of people
arrested. She was miraculously released, but her schoolmate,
arrested at the same time, was executed.
Her first scientific work, in Straub's laboratory, was on actin. She
tried, unsuccessfully, to show that actin had an intrinsic ATPase ac-
tivity, a fact that was demonstrated later by others. She then partly
purified an ATPase from erythrocytes: it was the first membrane
bound ATPase, but this was published in a practically unknown
journal. In the early 1950s, she became interested in protein biosyn-
thesis and hoped to obtain it in vitro. After an unsuccessful attempt
to reproduce the synthesis of cytochrome C, described in a publica-
tion from Frtiz Lipmann's laboratory that later proved to be a fake,
she tried to obtain the synthesis of amylase in various cell-free sys-
tems. She obtained an increase in amylase enzymatic activities but
when she managed to obtained some radioactive amino-acids, she
found no specific labeling of the protein. She concluded that the in-
crease in activity could correspond to the conversion of a precursor.
This work formed the substance of her doctoral thesis.
One of the consequences of the Communist dictatorship in
Hungary was that teaching of genetics was forbidden. Only the the-
ories of the Russian agronomist Trofim Lysenko were taught. He
rejected the science of genetics, believed in the heritability of ac-
quired characteristics, and claimed that their inheritance played a
major role in evolution. He considered Mendel's principles incom-
patible with dialectic materialism, which pleased Stalin. He not
only succeeded in ruining Soviet agriculture but also caused the
elimination of the best Soviet geneticists. It was during the Lysenko
affair, in 1948, that Agnes Ullmann first heard of Jacques Monod. A
friend of her had succeeded in smuggling a French newspaper
called Combat, directed by Albert Camus and dated September 19,
1948. In it was an article by a certain Dr. Monod entitled “The Vic-
tory of Lyssenko Has No Scientific Basis.” To her the article came as a
revelation and she decided on the spot that she would meet this Dr.
Monod one day, a wish that would come true 10 years later.
In 1956, Agnes actively participated in the revolution, when the
Hungarians hoped for a few days that they had freed themselves
from the Soviets. But on November 4, the Russian army invaded
the country to slash the revolution. Hundreds of people were
arrested and more than 1% of the population fled the country. By
the end of the year, Agnes was back at the bench and resumed
teaching, but times were hard because, once again, people were
living in constant fear.
118 In Memoriam / Research in Microbiology 170 (2019) 117e120
In 1957, having received an invitation to attend a Symposium of
the Society for Experimental Biology in London, she was lucky
enough to obtain a passport, and even a French visa. So, she left
for Paris and managed to meet the famous Dr. Monod who had
been fighting against Lysenko's theories. This was January 1958.
Agnes was an attractive 30 years old fair-haired woman.
When Monod asked her what she was doing in Paris, she boldly
told him that if he allowed her to work in his lab at Institut Pasteur,
she could extend her stay to six weeks. The next morning, she
started working with François Gros on the inhibition of protein syn-
thesis by chloramphenicol. Her collaboration with François Gros
developed into a close friendship, which would last all of her life.
One day, she confided to him that she wanted to leave Hungary
for good, come to France, and go on working forever in the labora-
tory. François Gros advised her to discuss this problem with Monod
but, since she did not dare to do it, he did it for her. The next day
Jacques Monod invited her for dinner at his home. After a few hours
during which she recounted what had happened during the revo-
lution, the unending menace of persecution under which Hungar-
ians were living, constantly fearing for their security, Jacques
Monod told her that he was willing to do everything in his power
to help her. When she asked him why, he simply answered: “It is
a matter of human dignity.”
Back in Budapest, taking advantage of the techniques she had
learnt from François Gros, she decided to study, together with her
husband, Tom Erdo €s, a former student of Szent-Gyo€ rgyi, the mech-
anism of action of streptomycin. Working with streptomycin-
sensitive, resistant and dependent strains of Mycobacterium fribur-
giensis, they were able to show, for the first time, that streptomycin
inhibits protein synthesis.
Meanwhile, Monod had not forgotten his promise. And he orga-
nized the escape of Agnes and her husband, aboard a trailer pulled
by the car of an “Austrian tourist”. “Our escape, she wrote, was
scheduled for a Saturday in June 1960. The trailer arrived as
planned and we crammed ourselves under a bathtub. At the border,
two customs officers searched every corner of the vehicle with a
flashlight for more than one hour. It was an absolute miracle that
they did not catch us. The officers finally left the trailer and, after
a few more controls, we were allowed to proceed.”
Once in Paris, Agnes went to work in Monod's laboratory, while
her husband Tom worked at a research institute outside of Paris.
Monod was then working on the regulation of the synthesis of pro-
teins involved in lactose metabolism in Escherichia coli. The operon
model was taking shape, and it involved the participation of a
repressor, preventing the synthesis of the proteins in the absence
of lactose. The first project given by Monod to Agnes was the isola-
tion of this repressor, then believed to be an RNA. Not surprisingly,
she was not successful, first because the repressor turned out to be
a protein, and, second, because she lacked a strain overproducing
the repressor, as did Wally Gilbert and Benno Müller-Hill, who iso-
lated it a few years later.
According to the model, the transcription of genes in an
operon starts at a DNA sequence called the promoter, overlapping
or close to another sequence, the operator, corresponding to the
repressor binding site. Therefore, if one could join structural
genes of one operon to the promoter/operator sequences of
another, this would result in the subordination of those structural
genes to the regulatory mechanism of the other operon. This is
what François Jacob and Agnes Ullmann succeeded to do, in the
end of 1963, by obtaining and analyzing mutants carrying dele-
tions that fused genes from the lactose operon to the promoter
of an operon involved in the synthesis of purines. These experi-
ments represented the first example of in vivo genetic engineer-
ing. Gene fusions were to become a major tool to study genetic
analysis and regulation.
Work with the lactose system uncovered seemingly contradic-
tory complementation results concerning lacZ, the gene encoding
b-galactosidase. Although there was strong genetic and biochem-
ical evidence that the gene was a single cistron, encoding a single
polypeptide chain, enzyme activity could be obtained in strains
containing one gene copy bearing a deletion at its promoter distal
end and one copy with a mutation at its promoter proximal end.
Exactly what would be expected if the gene consisted of two cis-
trons. Agnes had a major contribution in solving this apparent
contradiction. This unexpected complementation involved nonco-
valent association between peptides corresponding to different
fragments of the wild-type chain.
While this first example of intracistronic complementation,
then called u-complementation, had been observed using genes
deleted in the distal part, Agnes wondered whether other combina-
tions of deletion mutants would generate an active enzyme. And
she found that when extracts from strains carrying partial deletions
of the promoter proximal segment of lacZ were mixed with extracts
of b-galactosidase-negative mutants whose promoter-proximal
segments were intact, she recovered enzymatic activity. There
again, she concluded that a peptide encoded by the proximal part
of the gene can combine with the rest of the polypeptide to form
an active enzyme (so-called a-complementation). When the crystal
structure of b-galactosidase was published many years later, Agnes
was delighted to find that her original hypothesis based on
biochemical and genetic experiments had been fully confirmed.
Today, a-complementation has become the basis for the commonly
used “blue/white screen” to identify recombinant DNA: insertion of
foreign DNA into the plasmid-borne proximal part of lacZ results in
the abolition of a-complementation and, therefore, of the ability of
the recombinant cells to hydrolyze the dye XGal.
In 1961, once the historical paper on “Genetic regulatory mech-
anisms in the synthesis of proteins” had been published, Monod
became interested in another type of regulation, that which gov-
erns the activity of enzymes. How compounds bearing no structural
similarity with the enzyme substrate could activate or inhibit this
enzyme? This was to be the great period of “allostery”. There again,
Agnes took her part, working on rabbit muscle phosphorylase b, an
enzyme activated by 50 -AMP. She contributed in showing that in
the presence of 50 -AMP, phosphorylase b undergoes a reversible
conformational change that increases its capacity to bind certain li-
gands. This was to be the basis of the allosteric model: activators or
inhibitors, binding to a site different from the catalytic site, stabilize
the enzyme in a new conformation.
When Monod was appointed professor at the Colle ge de France,
in 1967, he asked Agnes to give a seminar. She chose to talk about a
cascade of covalent regulations occurring in the muscle and in
which 30 ,50 -cyclic adenosine monophosphate (cAMP) plays the
role of a second messenger which activates a cAMP-dependent pro-
tein kinase. When she read a paper by Makman and Sutherland1
describing the presence of cAMP in bacteria, she decided to look
for covalent modifications in E. coli triggered by cAMP. This, she
did not find, but instead she soon found evidence that cAMP was
an allosteric effector involved in the regulation of transcription.
From her results and those of others, it was found that, by binding
to CAP (the catabolite gene activator protein, its receptor protein),
cAMP exerts a positive regulation on the otherwise negatively regu-
lated lac operon. For 18 years, Agnes worked on the role of cAMP in
regulatory mechanisms in bacteria, and with her collaborators, -
because by then she had her own group - she published approxi-
mately 50 papers on the subject.
1
Makman R.S Sutherland EW (1965) Adenosine 30 ,50 -Phosphate in Escherichia
Coli J. Biol. Chem. 240:1309e14.
 In Memoriam / Research in Microbiology 170 (2019) 117e120
In the early 1980s, with the collaboration of Antoine Danchin,
Agnes started to study the enzyme producing cAMP, adenylate
cyclase. Danchin cloned the E. coli gene for this enzyme, cyaA,
and prepared a gene fusion with lacZ, encoding a hybrid protein
endowed with both enzymatic activities. Agnes, using a one-step
purification procedure that she had devised for b-galactosidase, pu-
rified this hybrid protein. This allowed them to perform a structure/
function study of adenylate cyclase.
Having spent many years on the role of cAMP in E. coli, she
decided to switch to a different area, i.e., study the role of this
nucleotide in pathogenesis.
Upon reviewing the relevant literature, she discovered reports
on the existence in Bordetella pertussis, the etiologic agent of
whooping cough, of an adenylate cyclase that exhibited two strik-
ing properties: (i) it was secreted into the extracellular space
(only intracellular adenylate cyclases were known at that time)
and (ii) it was activated by calmodulin, a eukaryotic protein. This
enzyme was soon demonstrated to act as a toxin able to invade an-
imal cells where, upon activation by calmodulin, it elicits an unreg-
ulated increase in cAMP, thereby disrupting normal cellular
functions. The study of this fascinating protein then became a major
endeavour, implying both Agnes' group in close collaboration with
the team of A. Danchin as well as several external collaborations,
some within a Human Frontier Science Program (HFSP) entitled
“Molecular and Cellular Mechanisms Underlying Regulation of
Virulence Factors of B. pertussis” coordinated by her. Analysis of
the protein, CyaA, and its gene revealed that the amino-terminal
end of the protein bears the calmodulin activated catalytic activity
whereas the carboxy-terminal exhibits strong homology to the E.
coli hemolysin. The latter is responsible for the binding of the toxin
to eukaryotic cells, and for its internalization. Studies of the genetic
organization of the cya locus revealed the presence, downstream to
the adenylate cyclase structural gene (cyaA), of three additional
genes (cyaB, cyaD, and cyaE) that build a type I ATP-Binding
Cassette (ABC) secretion machinery dedicated to the secretion of
the toxin. Agnes later got interested in deciphering the transcrip-
tional regulation of the cya operon by a two-component regulatory
system (BvgA/BvgS) that controls the expression of the virulent
genes of B. pertussis.
An important development of this study has been the use of
adenylate cyclase as a vector to deliver specific epitopes into anti-
gen presenting cells. Indeed, several permissive sites were found
in the gene where the insertion of a few codons would not impair
any of the properties of the protein. Further work demonstrated
that such recombinant toxins represented an attractive non-
replicative vector for antigen delivery and could be used for the
development of protective and therapeutic vaccines. After her
mandatory retirement in 1995, Agnes continued to closely follow
the development of this project that lead to the design of two re-
combinant vaccines that went into phase I/II clinical trials in cancer
immunotherapy.
A mere examination of her research activity fails to give full
credit to the extraordinary personality of Agnes Ullmann. Her pas-
sion for science was testified, for instance by her assiduity to sem-
inars, to which she always participated actively as long as she could
come to Institut Pasteur, until the very end of 2016. Despite severe
health problems - she was progressively losing her sight due to a
Macular Degeneration and suffered from hard back pain and had
difficulty to breathe - Agnes was still coming several days a week
to Institut Pasteur where she had a small office. And she was
constantly reminding young - or even less young - researchers to
attend seminars, and to those who would argue that they could
not as they had experiments ongoing, she would answer: « your
119
experiments are not more important than that of others ». Together
with the regretted Ce
cile Wandersman, they were for decades the
pillars of the “Journal club de Microbiologie”, where many of the
young scientists and postdocs from the institute had a chance to
calibrate their critical mind on theirs.
She was also devoted to teaching and organizing courses at the
institute as well as in Greece, Brazil or India. She was always
engaged in the support of brilliant young scientists, for instance
by presiding from its beginning, in 1979, the scientific committee
of a “Jacques Monod Foundation” that gave each year one or two
prizes to young scientists. By the way, presiding this committee
was one of the many ways in which she paid homage to the mem-
ory of Jacques Monod, who died in 1976, and for whom she had a
profound devotion.
This devotion, she expressed in many ways. Soon after Monod's
death, she co-edited with Andre
Lwoff “Selected papers in molecular
biology by Jacques Monod, as well as, “Origins of Molecular Biology. A
tribute to Jacques Monod” ” with accounts from many scientists who
had worked with or close to him both at Academic Press Inc. (1978
and 1979, respectively). In 2003, she obtained the publication of a
revised edition of the second of these books by ASM Press. A
more personal expression of this devotion was the care she took
of Monod's Airedale that she brought with her to the lab
everyday… even though dogs were, in principle, not allowed in
the institute.
During her career Agnes also organized numerous scientific
meetings including a prestigious celebration of the Centennial of
the Institut Pasteur in 1987, and another one for the 100th anniver-
sary of Louis Pasteur death in 1995.
From 1982 to 1995, under the directorship of Raymond
Dedonder and then of Maxime Schwartz, she was part of the direc-
tion of Institut Pasteur, in charge of the applications of research.
One of her feat in this function has been the setting up of a “com-
mando”, headed by Simon Wain-Hobson, to sequence the HIV
genome. They came first, before the American competitors, and
this was important for the patents in this domain.
Her devotion for science included a determination to avoid
excess spending of research money. She was not of the kind to
travel business class, take taxis when not obligatory, ask for reim-
bursement of expensive meals or hotels, etc.
One of her most obvious qualities was courage. Courage during
her youth, when she opposed the soviet regime in her country.
Courage when she escaped from Hungary. Even courage during
the last months of her life, when she was alone, almost blind,
with great difficulties to breathe, and who, when you visited her,
only asked questions about you and your family. Another hallmark
of her personality was a genuine curiosity she kept intact up to her
very last days: “what's new, in the lab, in “Pasteur”, in science, etc.
?” were her favorite words when one would call or visit her. She
enjoyed discussing about recently published articles or ongoing ex-
periments. And despite being confined in her apartment she was
still eagerly interested in what was going on in the world, in her
countries e of origin and of adoption e and in her true “home”, l’In-
stitut Pasteur.
Agnes Ullmann was both a member of the CNRS (1962e1992)
and of Institut Pasteur (1978e1996). She published 180 scientific
articles, edited or coedited 6 books, and cosigned 5 patents. Aside
from several French decorations, including the Legion of Honor,
she received the Robert Koch medal (Germany), was Doctor Hono-
ris Causa of the Sapienza University in Rome, was a member of
EMBO, of the Hungarian Science Academy and of the European
Microbiology Academy. Finally, in 1966, she had obtained French
citizenship, something she was very proud of.
120 In Memoriam / Research in Microbiology 170 (2019) 117e120

Agnes was a woman of great culture in the authentic tradi- Daniel Ladant
tion of the “Mitteleuropa”. Over the last seven decades, she Unit

e de Biochimie des Interactions Macromol
eculaires, Institut
enjoyed working and interacting with many great scientists in Pasteur, CNRS, Paris, France
many countries. She had witnessed and contributed to the birth
Didier Mazel*
and development of modern biology. She also liked to recall that
Unit

e de Plasticit

e du G

enome Bact

erien, Institut Pasteur, CNRS, Paris,
she had known an era “before DNA”, a time when to perform an
France
experiment, she had first to purify ATP from rabbit muscle, and
she was kindly making fun of the present day “kit generation”. Maxime Schwartz
She died on February 25, 2019. Direction de la Communication, Institut Pasteur, Paris, France
It has been a privilege for us to have been students, colleagues
and friends of this Great Lady. *
Corresponding author.
E-mail address: didier.mazel@pasteur.fr (D. Mazel).
Conflict of interest
10 April 2019
None. Available online 15 May 2019