Charles C. Richardson

Last updated

Charles C. Richardson
Born(1935-05-07)May 7, 1935
Wilson, NC, United States
Alma mater Duke Medical School
Duke University
SpouseIngrid Hanssum (1961; 2 children)
Scientific career
Fields Molecular biology
Institutions Harvard University

Charles Clifton Richardson (born May 7, 1935) is an American biochemist and professor at Harvard University. Richardson received his undergraduate education at Duke University, where he majored in medicine. He received his M.D. at Duke Medical Schoo l in 1960. Richardson works as a professor at Harvard Medical School, and he served as editor/associate editor of the Annual Review of Biochemistry from 1972 to 2003. [1] Richardson received the American Chemical Society Award in Biological Chemistry in 1968, as well as numerous other accolades. [2]

Contents

Early life and education

Charles Richardson was born on May 7, 1935, in Wilson, North Carolina. [1] His father, Barney Clifton Richardson, was an accountant at a local automobile dealership. His mother, Elizabeth Barefoot, was a housewife. At 11 years old, Richardson and his family moved to Columbia, South Carolina. Richardson graduated from Dreher High School and received a full scholarship to Duke University in 1953. Without completing a bachelor's degree, Richardson enrolled in Duke Medical School in 1956. In 1959, Richardson completed a Bachelor of Science degree in medicine from Duke through the National Institutes of Health (NIH) United States Public Health Service Post-Sophomore Research Fellowship. Richardson graduated from Duke Medical School and began residency at Duke University Hospital in 1960. On July 29, 1961, Richardson married Ingrid Hanssum at the Gothic Duke Chapel. They have two children. [1]

Career and research

In 1961, Richardson obtained a Public Health Service fellowship under Arthur Kornberg in his biochemistry laboratory at Stanford Medical School. As a result, Richardson and Ingrid Hanssum moved to Palo Alto. In Kornberg's lab, Richardson focused on improving the purification technique of DNA polymerase from E. coli . In Kornberg's lab, Richardson worked alongside Paul Berg, Reiji and Tsunko Okazaki, and several others. In 1964, Richardson left Kornberg's lab and began a faculty position at Harvard Medical School, where he was promoted to tenure in 1967. Richardson served as chairman of the department of biological chemistry from 1978 to 1987. Additionally, Richardson served as editor or associate editor of the Annual Review of Biochemistry from 1972 to 2003. As of 2020, Richardson continues his position as professor at Harvard Medical School. [1] Richardson taught four doctoral students: Dennis M. Livingston, David N. Frick, Richard D. Colodner, and Paul L. Modrich. [3]

Throughout Richardson's career, Richardson used bacteriophages in order to investigate DNA replication. Richardson discovered and researched several enzymes throughout his career: E. coli exonuclease III [4] in 1964, T4 DNA ligase [5] in 1967, T7 DNA polymerase [6] in 1971, E. coli exonuclease VII [7] [8] in 1974, E. coli DNA polymerase III [9] [10] in 1975, T4 polynucleotide kinase [11] in 1981, T7 DNA primase [12] [13] in the late 1980s and early 1990s, and T7 DNA helicase [14] in 2004. Richardson used these enzymes to further analyze DNA, develop sequencing reagents, and characterize the mechanisms of DNA replication. [15]

Richardson's most highly-cited accomplishment was made while working with bacteriophage T7 RNA polymerase in 1985. Richardson used the T7 RNA polymerase/promoter system to control the expression of a phage T7 gene 5 protein (gp5), which is a subunit of T7 DNA polymerase. By combining the specificity of T7 RNA polymerase for its own promoters with rifampicin's ability to selectively inhibit the host RNA polymerase, Richardson established a method to exclusively express genes, specifically the phage T7 gene 5 protein, under the control of the T7 RNA polymerase promoter. During this process, Richardson constructed a T7 phage with deletions in gene 1 that propagate in E. coli cells expressing T7 RNA polymerase. Richardson proposed the T7 RNA polymerase/promoter system as an "attractive alternative" to the mini- or maxicell. [16]

A couple years later, Richardson researched a self-made DNA polymerase for potential use in DNA sequencing. This highly processive DNA polymerase was composed of an 84-kDa T7 gene 5 protein and 12-kDa E. coli thioredoxin at a one-to-one stoichiometric ratio. [17] In his study, Richardson demonstrated that this modified DNA polymerase would be ideal for DNA sequencing by the chain-termination method. Richardson based this finding off of three main factors: high processivity and lack of associated exonuclease activity, ability to use low concentrations of radioactive nucleotides for preparation of DNA probes, and lack of background pause sites and uniform distribution of dideoxy-terminated fragments. [18]

In 1998, Richardson examined the crystal structure of a bacteriophage T7 DNA replication complex at 2.2 Å resolution. Before imaging, Richardson complexed the T7 bacteriophage DNA polymerase with a primer-template and a nucleoside triphosphate in the polymerase active site. Through analysis of the crystal structure, Richardson determined how the replication complex selects nucleotides in a template-directed manner. Furthermore, Richardson established an understanding of the basis for phosphoryl transfer by related polymerases with metal. [19]

More recently in 2011, Richardson developed a single-molecule assay to measure the activity of the replisome with fluorescently-labeled DNA polymerases. Richardson then used this assay to quantify the process of polymerase exchange. Richardson determined that soluble polymerases are recruited to an actively synthesizing replisome, which leads to a polymerase exchange event between the excess polymerases and the synthesizing polymerase after about 50 seconds. This supports the belief that replisomes are highly dynamic complexes. [20]

Awards and honors

Memberships

Related Research Articles

<span class="mw-page-title-main">DNA ligase</span> Class of enzymes

DNA ligase is a type of enzyme that facilitates the joining of DNA strands together by catalyzing the formation of a phosphodiester bond. It plays a role in repairing single-strand breaks in duplex DNA in living organisms, but some forms may specifically repair double-strand breaks. Single-strand breaks are repaired by DNA ligase using the complementary strand of the double helix as a template, with DNA ligase creating the final phosphodiester bond to fully repair the DNA.

<span class="mw-page-title-main">DNA polymerase</span> Form of DNA replication

A DNA polymerase is a member of a family of enzymes that catalyze the synthesis of DNA molecules from nucleoside triphosphates, the molecular precursors of DNA. These enzymes are essential for DNA replication and usually work in groups to create two identical DNA duplexes from a single original DNA duplex. During this process, DNA polymerase "reads" the existing DNA strands to create two new strands that match the existing ones. These enzymes catalyze the chemical reaction

<span class="mw-page-title-main">DNA polymerase I</span> Family of enzymes

DNA polymerase I is an enzyme that participates in the process of prokaryotic DNA replication. Discovered by Arthur Kornberg in 1956, it was the first known DNA polymerase. It was initially characterized in E. coli and is ubiquitous in prokaryotes. In E. coli and many other bacteria, the gene that encodes Pol I is known as polA. The E. coli Pol I enzyme is composed of 928 amino acids, and is an example of a processive enzyme — it can sequentially catalyze multiple polymerisation steps without releasing the single-stranded template. The physiological function of Pol I is mainly to support repair of damaged DNA, but it also contributes to connecting Okazaki fragments by deleting RNA primers and replacing the ribonucleotides with DNA.

dnaQ is the gene encoding the ε subunit of DNA polymerase III in Escherichia coli. The ε subunit is one of three core proteins in the DNA polymerase complex. It functions as a 3’→5’ DNA directed proofreading exonuclease that removes incorrectly incorporated bases during replication. dnaQ may also be referred to as mutD.

In molecular biology and biochemistry, processivity is an enzyme's ability to catalyze "consecutive reactions without releasing its substrate".

Site-directed mutagenesis is a molecular biology method that is used to make specific and intentional mutating changes to the DNA sequence of a gene and any gene products. Also called site-specific mutagenesis or oligonucleotide-directed mutagenesis, it is used for investigating the structure and biological activity of DNA, RNA, and protein molecules, and for protein engineering.

In molecular biology, endonucleases are enzymes that cleave the phosphodiester bond within a polynucleotide chain. Some, such as deoxyribonuclease I, cut DNA relatively nonspecifically, while many, typically called restriction endonucleases or restriction enzymes, cleave only at very specific nucleotide sequences. Endonucleases differ from exonucleases, which cleave the ends of recognition sequences instead of the middle (endo) portion. Some enzymes known as "exo-endonucleases", however, are not limited to either nuclease function, displaying qualities that are both endo- and exo-like. Evidence suggests that endonuclease activity experiences a lag compared to exonuclease activity.

<span class="mw-page-title-main">Exonuclease</span> Class of enzymes; type of nuclease

Exonucleases are enzymes that work by cleaving nucleotides one at a time from the end (exo) of a polynucleotide chain. A hydrolyzing reaction that breaks phosphodiester bonds at either the 3′ or the 5′ end occurs. Its close relative is the endonuclease, which cleaves phosphodiester bonds in the middle (endo) of a polynucleotide chain. Eukaryotes and prokaryotes have three types of exonucleases involved in the normal turnover of mRNA: 5′ to 3′ exonuclease (Xrn1), which is a dependent decapping protein; 3′ to 5′ exonuclease, an independent protein; and poly(A)-specific 3′ to 5′ exonuclease.

<span class="mw-page-title-main">DNA polymerase II</span>

DNA polymerase II is a prokaryotic DNA-dependent DNA polymerase encoded by the PolB gene.

Marlene Belfort is an American biochemist known for her research on the factors that interrupt genes and proteins. She is a fellow of the American Academy of Arts and Sciences and has been admitted to the United States National Academy of Sciences.

<span class="mw-page-title-main">Roger D. Kornberg</span> American biochemist and professor of structural biology

Roger David Kornberg is an American biochemist and professor of structural biology at Stanford University School of Medicine. Kornberg was awarded the Nobel Prize in Chemistry in 2006 for his studies of the process by which genetic information from DNA is copied to RNA, "the molecular basis of eukaryotic transcription."

Deoxyribonuclease IV (phage-T4-induced) is catalyzes the degradation nucleotides in DsDNA by attacking the 5'-terminal end.

<span class="mw-page-title-main">Thomas B. Kornberg</span> American biochemist (born 1948)

Thomas Bill Kornberg is an American biochemist who was the first person to purify and characterise DNA polymerase II and DNA polymerase III. He is currently a professor of biochemistry and biophysics at the University of California, San Francisco, and is working on Drosophila melanogaster development.

<span class="mw-page-title-main">T7 DNA polymerase</span>

T7 DNA polymerase is an enzyme used during the DNA replication of the T7 bacteriophage. During this process, the DNA polymerase “reads” existing DNA strands and creates two new strands that match the existing ones. The T7 DNA polymerase requires a host factor, E. coli thioredoxin, in order to carry out its function. This helps stabilize the binding of the necessary protein to the primer-template to improve processivity by more than 100-fold, which is a feature unique to this enzyme. It is a member of the Family A DNA polymerases, which include E. coli DNA polymerase I and Taq DNA polymerase.

PstI is a type II restriction endonuclease isolated from the Gram negative species, Providencia stuartii.

The history of genetics can be represented on a timeline of events from the earliest work in the 1850s, to the DNA era starting in the 1940s, and the genomics era beginning in the 1970s.

The Phage 21 S Family is a member of the Holin Superfamily II.

Mary P. Edmonds was an American biochemist who made key discoveries regarding the processing of messenger RNA (mRNA). She spent most of her career at the University of Pittsburgh.

Sylvy Kornberg née Sylvia Ruth Levy (1917–1986) was an American biochemist who carried out research on DNA replication and polyphosphate synthesis. She discovered and characterized polyphosphate kinase (PPK), an enzyme that helps build long chains of phosphate groups called polyphosphate (PolyP) that play a variety of metabolic and regulatory functions. She worked closely with her husband and research partner, Arthur Kornberg, contributing greatly to the characterization of DNA polymerization that earned him the 1959 Nobel Prize in Physiology or Medicine.

<span class="mw-page-title-main">George Stark</span>

George Stark is an American chemist and biochemist. His research interests include protein and enzyme function and modification, interferons and cytokines, signal transduction, and gene expression.

References

  1. 1 2 3 4 5 Richardson, Charles C. (June 2, 2015). "It Seems Like Only Yesterday". Annual Review of Biochemistry. 84 (1): 1–34. doi: 10.1146/annurev-biochem-060614-033850 . ISSN   0066-4154. PMID   26034887.
  2. 1 2 3 4 5 6 7 8 9 10 11 12 13 Richardson, Charles. "Curriculum Vitae". Charles C. Richardson Laboratory. Retrieved March 8, 2020.
  3. ↑ "Chemistry Tree - Charles C. Richardson Family Tree". academictree.org. Retrieved April 19, 2020.
  4. ↑ Richardson, Charles C.; Lehman, I. R.; Kornberg, Arthur (January 1, 1964). "A Deoxyribonucleic Acid Phosphatase-Exonuclease from Escherichia coli II. Characterization of the Exonuclease Activity". Journal of Biological Chemistry. 239 (1): 251–258. doi: 10.1016/S0021-9258(18)51775-0 . ISSN   0021-9258. PMID   14114851.
  5. ↑ Fareed, G C; Richardson, C C (1967). "Enzymatic breakage and joining of deoxyribonucleic acid. II. The structural gene for polynucleotide ligase in bacteriophage T4". Proceedings of the National Academy of Sciences of the United States of America. 58 (2): 665–672. Bibcode:1967PNAS...58..665F. doi: 10.1073/pnas.58.2.665 . ISSN   0027-8424. PMC   335686 . PMID   5234326.
  6. ↑ Grippo, Pasquale; Richardson, Charles C. (November 25, 1971). "Deoxyribonucleic Acid Polymerase of Bacteriophage T7". Journal of Biological Chemistry. 246 (22): 6867–6873. doi: 10.1016/S0021-9258(19)45926-7 . ISSN   0021-9258. PMID   4942327.
  7. ↑ Chase, John W.; Richardson, Charles C. (July 25, 1974). "Exonuclease VII of Escherichia coli Purification and Properties". Journal of Biological Chemistry. 249 (14): 4545–4552. doi: 10.1016/S0021-9258(19)42453-8 . ISSN   0021-9258. PMID   4602029.
  8. ↑ Chase, John W.; Richardson, Charles C. (July 25, 1974). "Exonuclease VII of Escherichia coli Mechanism of Action". Journal of Biological Chemistry. 249 (14): 4553–4561. doi: 10.1016/S0021-9258(19)42454-X . ISSN   0021-9258. PMID   4602030.
  9. ↑ Livingston, D. M.; Hinkle, D. C.; Richardson, C. C. (January 25, 1975). "Deoxyribonucleic acid polymerase III of Escherichia coli. Purification and properties". Journal of Biological Chemistry. 250 (2): 461–469. doi: 10.1016/S0021-9258(19)41920-0 . ISSN   0021-9258. PMID   1089643.
  10. ↑ Livingston, D. M.; Richardson, C. C. (January 25, 1975). "Deoxyribonucleic acid polymerase III of Escherichia coli. Characterization of associated exonuclease activities". Journal of Biological Chemistry. 250 (2): 470–478. doi: 10.1016/S0021-9258(19)41921-2 . ISSN   0021-9258. PMID   163228.
  11. ↑ Richardson, Charles C. (January 1, 1981), Boyer, Paul D. (ed.), 16 Bacteriophage T4 Polynucleotide Kinase, The Enzymes, vol. 14, Academic Press, pp. 299–314, doi:10.1016/S1874-6047(08)60342-X, ISBN   9780121227142 , retrieved April 17, 2020
  12. ↑ Bernstein, J. A.; Richardson, C. C. (August 5, 1989). "Characterization of the helicase and primase activities of the 63-kDa component of the bacteriophage T7 gene 4 protein". Journal of Biological Chemistry. 264 (22): 13066–13073. doi: 10.1016/S0021-9258(18)51596-9 . ISSN   0021-9258. PMID   2546945.
  13. ↑ Mendelman, L. V.; Notarnicola, S. M.; Richardson, C. C. (December 25, 1993). "Evidence for distinct primase and helicase domains in the 63-kDa gene 4 protein of bacteriophage T7. Characterization of nucleotide binding site mutant". Journal of Biological Chemistry. 268 (36): 27208–27213. doi: 10.1016/S0021-9258(19)74239-2 . ISSN   0021-9258. PMID   8262962.
  14. ↑ Crampton, Donald J.; Richardson, Charles C. (January 1, 2003). "Bacteriophage T7 gene 4 protein: A hexameric DNA helicase". In Hackney, David D.; Tamanoi, Fuyuhiko (eds.). Energy Coupling and Molecular Motors. The Enzymes. Energy Coupling and Molecular Motors. Vol. 23. Academic Press. pp. 277–302. doi:10.1016/S1874-6047(04)80007-6. ISBN   9780121227241 . Retrieved April 17, 2020.
  15. ↑ Kresge, Nicole; Simoni, Robert D.; Hill, Robert L. (July 13, 2007). "DNA Replication in Bacteriophage: the Work of Charles C. Richardson". Journal of Biological Chemistry. 282 (28): e22. doi: 10.1016/S0021-9258(19)78070-3 . ISSN   0021-9258.
  16. ↑ Tabor, S.; Richardson, C. C. (February 1, 1985). "A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes". Proceedings of the National Academy of Sciences. 82 (4): 1074–1078. Bibcode:1985PNAS...82.1074T. doi: 10.1073/pnas.82.4.1074 . ISSN   0027-8424. PMC   397196 . PMID   3156376.
  17. ↑ Mark, D. F.; Richardson, C. C. (March 1, 1976). "Escherichia coli thioredoxin: a subunit of bacteriophage T7 DNA polymerase". Proceedings of the National Academy of Sciences. 73 (3): 780–784. Bibcode:1976PNAS...73..780M. doi: 10.1073/pnas.73.3.780 . ISSN   0027-8424. PMC   336002 . PMID   768986.
  18. ↑ Tabor, S.; Richardson, C. C. (July 1, 1987). "DNA sequence analysis with a modified bacteriophage T7 DNA polymerase". Proceedings of the National Academy of Sciences. 84 (14): 4767–4771. Bibcode:1987PNAS...84.4767T. doi: 10.1073/pnas.84.14.4767 . ISSN   0027-8424. PMC   305186 . PMID   3474623.
  19. ↑ Doublié, Sylvie; Tabor, Stanley; Long, Alexander M.; Richardson, Charles C.; Ellenberger, Tom (1998). "Crystal structure of a bacteriophage T7 DNA replication complex at 2.2 Å resolution". Nature. 391 (6664): 251–258. Bibcode:1998Natur.391..251D. doi:10.1038/34593. ISSN   1476-4687. PMID   9440688. S2CID   4384241.
  20. ↑ Loparo, Joseph J.; Kulczyk, Arkadiusz W.; Richardson, Charles C.; van Oijen, Antoine M. (January 18, 2011). "Simultaneous single-molecule measurements of phage T7 replisome composition and function reveal the mechanism of polymerase exchange". Proceedings of the National Academy of Sciences. 108 (9): 3584–3589. doi: 10.1073/pnas.1018824108 . ISSN   0027-8424. PMC   3048139 . PMID   21245349.

Further reading

This page is based on this Wikipedia article
Text is available under the CC BY-SA 4.0 license; additional terms may apply.
Images, videos and audio are available under their respective licenses.