40 Years with Bacteriophage Ø29

Annu. Rev. Microbiol. 2007.61:1-22. Downloaded from arjournals.annualreviews.org ANRV322-MI61-01 ARI 4 September 2007 23:12 by CENTRO BIOLOGIA MOLECULAR on 02/11/08. For personal use only. ANRV322-MI61-01 ARI 4 September 2007 23:12 40 Years with Bacteriophage ø29 Margarita Salas Instituto de Biologıa´ Molecular “Eladio Vinuela”˜ (CSIC), Centro de Biologıa´ Molecular “Severo Ochoa” (CSIC–UAM), Universidad Autonoma,´ Cantoblanco, 28049 Madrid, Spain; email: [email protected] Annu. Rev. Microbiol. 2007. 61:1–22 Key Words First published online as a Review in Advance on DNA polymerase, protein-priming, replication, terminal protein, April 18, 2007 transcription The Annual Review of Microbiology is online at micro.annualreviews.org Abstract This article’s doi: I have dedicated the past 46 years of my life to science and I expect 10.1146/annurev.micro.61.080706.093415 to be active in research for many more years. I have been lucky in by CENTRO BIOLOGIA MOLECULAR on 02/11/08. For personal use only. Copyright c 2007 by Annual Reviews. my professional life. During my postdoctoral years I discovered two ! All rights reserved Annu. Rev. Microbiol. 2007.61:1-22. Downloaded from arjournals.annualreviews.org proteins that I showed to be involved in the initiation of protein 0066-4227/07/1013-0001$20.00 synthesis. Working with bacteriophage ø29 for the past 40 years, we have made many interesting findings. Among them is the discovery of a protein covalently linked to the 5! ends of ø29 DNA that we later showed to be the primer for the initiation of ø29 DNA replication. Also, the finding of the ø29 DNA polymerase with its properties of high processivity, strand displacement, and high fidelity has been very rewarding. The ø29 DNA polymerase has become the ideal en- zyme for DNA amplification, both rolling circle and whole-genome amplification. I also am happy because I have worked with many brilliant students and collaborators over the years, most of whom have become excellent scientists. 1 ANRV322-MI61-01 ARI 4 September 2007 23:12 beginning of the Spanish Civil War, and in Contents 1937 they left Madrid and went to Canero, where my grandparents had a country house. CHILDHOOD AND YOUTH . 2 They had three children, my brother, Jose,´ FROM ORGANIC CHEMISTRY who is one year older than me, myself, and TO BIOCHEMISTRY ........... 3 my sister, Marıa´ Luisa (Marisa), who is two THREE YEARS AT NEW YORK years my junior. My parents stayed in Canero UNIVERSITY................... 5 until the end of the Civil War, in 1939. Then, BACK TO SPAIN: PHAGE ø29 AS A they moved to Gijon,´ also located on the north MODEL SYSTEM .............. 7 coast of Asturias, where my father set up a psy- EARLY YEARS IN MADRID . 8 chiatric clinic. My mother, who had to stop CONTROL OF ø29 DNA her career as a school teacher, managed the TRANSCRIPTION . 11 clinic. The house where the clinic was estab- PROTEIN-PRIMED ø29 DNA lished had three floors and one basement. The REPLICATION . 13 patients occupied the second and third floors, A Sliding-Back Mechanism to we lived on the first floor, and the basement Initiate TP-Primed DNA was used for general services such as kitchen, Replication.................... 13 laundry, and so on. We liked spending time Structural-Functional Studies on with the patients and listening to their sto- the ø29 DNA Polymerase and ries. The house was surrounded by a garden Terminal Protein . 14 with a tennis court where we enjoyed playing Viral Proteins p6 and p5 Essential tennis (which I still like but no longer play). for ø29 DNA Replication . 15 As my father was fond of classical music and In Vitro Amplification of opera, during the early years of my life I lis- ø29 DNA . 16 tened to music at home very often, which led Membrane Proteins Involved in to my appreciation of classical music. ø29 DNA Replication: Proteins When I was four years old I began attend- p1 and p16.7 . 17 ing a Catholic school run by nuns, which was Other Viral Proteins Involved in usual in Spain during the 1940s. There, I re- ø29 DNA Replication: Proteins ceived a good classical education, based in sci- p17 and p56 . 17 ence and humanities, which included Latin Phage-Host Interactions in ø29 and Greek. I studied French and only a few Development . 18 years of English. I received the Baccalaureate OTHER COLLABORATIONS . 18 title in 1954. My parents were very clear that FINAL REMARKS . 19 by CENTRO BIOLOGIA MOLECULAR on 02/11/08. For personal use only. the three of us should attend university, which Annu. Rev. Microbiol. 2007.61:1-22. Downloaded from arjournals.annualreviews.org was not common for women. My father al- ways told us that the only inheritance he was going to leave us was a university career. That CHILDHOOD AND YOUTH was, indeed, the best legacy he left us. In or- I was born in Canero, Spain, on November der to attend the university, it was mandatory 30, 1938. Canero is a small village located in to spend one year doing the so-called pre- Asturias, on the north coast of Spain, close to university studies. It was then time to choose Luarca, where the Nobel Prize winner Severo whether I wanted to follow a scientific or a Ochoa was born in 1905. humanistic career. My decision was clear: I My father was a physician who specialized wanted a scientific career. When I finished in psychiatry and neurology, and my mother, my preuniversity studies in science, I had to who is still alive, was a school teacher. My par- choose from the scientific careers offered a ents married in Madrid in 1936, just after the field in which I would specialize. I liked both 2 Salas ANRV322-MI61-01 ARI 4 September 2007 23:12 medicine and chemistry, the former likely in- One day that summer, Severo and Carmen fluenced by the fact that my father was a physi- came to visit us and had lunch at our house. I cian. Because I could only study chemistry remember we ate an excellent paella. On that at the University of Oviedo, which was the occasion I had the privilege of meeting Severo university closest to Gijon,´ I decided, with and talking with him about what I was doing the agreement of my parents, to attend the and about my thoughts regarding my future. Complutense University of Madrid (UCM) to The next day Severo was giving a conference start the first year that was common for both on his work in the nearby city of Oviedo and degrees. This allowed me one more year to de- asked my father and me to accompany him. cide. After I completed my first year I decided The three of us went to Oviedo in my father’s to continue to study chemistry. I think that car. Severo was a brilliant speaker and I be- was a good decision because I soon found my- came fascinated by his scientific work. I re- self fascinated by the laboratory work that we member he spoke about fatty acid metabolism. carried out. During my third year, especially, At that time I didn’t know much about bio- I spent many hours doing experimental work chemistry because I had not yet studied it in organic chemistry, which I liked so much at the university; Ochoa promised to send that I thought I would become a researcher in me a biochemistry book. I was excited when organic chemistry. one month later I received the book General Biochemistry, by Joseph S. Fruton and Sofia Simmonds, a classic, dedicated by Severo FROM ORGANIC CHEMISTRY Ochoa. When I was about to finish univer- TO BIOCHEMISTRY sity I had made up my mind to dedicate I spent the summer holidays during my years myself to biochemistry, and I told Severo at UCM in Gijon,´ where my parents lived. about my decision. He advised me to do my In the summer of 1958, when I had finished PhD thesis in Madrid with an excellent bio- my third year of chemistry, I was lucky to chemist, Alberto Sols, who had trained at the meet Severo Ochoa. Severo had left Spain Washington University School of Medicine in 1936, when the Spanish Civil War be- in St. Louis in the laboratory of Carl and gan, because he wanted to continue his re- Gerty Cori. Once I finished my PhD thesis, I search. After staying in Germany and Eng- could go to Severo’s lab in New York for post- land, in 1940 he went to the United States, doctoral training. Ochoa wrote me a refer- first to St. Louis, Missouri, to the Washington ence letter for Alberto Sols, who accepted me University School of Medicine, and then to right away, even though I was a woman, since New York, to the New York University Med- he could not refuse something requested by by CENTRO BIOLOGIA MOLECULAR on 02/11/08. For personal use only. ical Center. For some time he refused to visit Severo Ochoa, who by 1960 had already won Annu. Rev. Microbiol. 2007.61:1-22. Downloaded from arjournals.annualreviews.org Spain, even for vacation, but in 1958 he spent the Nobel Prize. I spent the next four years one month of summer in Asturias, half the working on my PhD thesis with Alberto Sols, time in Luarca, his hometown, and the other working on carbohydrate metabolism, specifi- half in Gijon,´ where his wife, Carmen, was cally on the glucosephosphate isomerase from born. Severo and my father were good friends yeast, which converts glucose-6-phosphate (they had studied medicine together at UCM, into fructose-6-phosphate.

Load more