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Discovery of bisulfite-mediated cytosine conversion to uracil, the key reaction for DNA methylation analysis--a personal account.

Hayatsu H - Proc. Jpn. Acad., Ser. B, Phys. Biol. Sci. (2008)

Bottom Line: At the same time, Shapiro and his coworkers in New York University found the same reaction independently.We also reported that 5-methylcytosine was deaminated by bisulfite only very slowly.The author's recent work that has resulted in an improvement of the procedure of analysis by use of a newly devised high concentration bisulfite solution is also described.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Pharmaceutical Sciences, Okayama University, Okayama, Japan. hayatsuh@cc.okayama-u.ac.jp

ABSTRACT
Methylation at position 5 of cytosine in DNA is being intensively studied in many areas of biological sciences, as the methylation is intimately associated with the control of gene functions. The principal analytical method for determining the sites of 5-methylcytosine in genome at the sequence level involves bisulfite modification of DNA. The utility of this chemical treatment is based on the property of bisulfite to selectively deaminate cytosine residues. The bisulfite-mediated cytosine deamination was discovered in 1970 by us in the University of Tokyo. At the same time, Shapiro and his coworkers in New York University found the same reaction independently. We also reported that 5-methylcytosine was deaminated by bisulfite only very slowly. These findings were later utilized by a group of Australian scientists to devise a means to analyze 5-methylcytosine in DNA; thus, a method called 'bisulfite genomic sequencing' was invented by these researchers in 1992. This review describes the author's reflection of the discovery of bisulfite reactions with pyrimidine bases. The author's recent work that has resulted in an improvement of the procedure of analysis by use of a newly devised high concentration bisulfite solution is also described.

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Bisulfite-mediated deamination of cytosine.
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f5-321: Bisulfite-mediated deamination of cytosine.

Mentions: These observations were all surprising. Sodium bisulfite is a common laboratory reagent. It was unbelievable that nobody had noticed these reactions in the past. We completed piling up necessary data within several months, wrote a short report, and submitted it in November, 1969, to J. Am. Chem. Soc. as a Communication. It was smoothly accepted, and in the end of that year, a 3-page proof-prints came in. Attached there was a message, saying that a similar study from another laboratory was presently in press and that the corrected proof should be returned as quickly as possible. Thus, we sent it back immediately, and kept watching the journal. A Communication by Shapiro and coworkers appeared in the January 28th issue of the journal and ours in the next issue February 11, 1970.2),3) Both had a diagram of the reaction scheme as shown here in Fig. 5. Their submission was done in October, a month earlier than ours. We were lucky that the journal did not turn down our paper. It was in fact quite fair to publish these almost identical contents from two independent groups in a journal virtually simultaneously.


Discovery of bisulfite-mediated cytosine conversion to uracil, the key reaction for DNA methylation analysis--a personal account.

Hayatsu H - Proc. Jpn. Acad., Ser. B, Phys. Biol. Sci. (2008)

Bisulfite-mediated deamination of cytosine.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3722019&req=5

f5-321: Bisulfite-mediated deamination of cytosine.
Mentions: These observations were all surprising. Sodium bisulfite is a common laboratory reagent. It was unbelievable that nobody had noticed these reactions in the past. We completed piling up necessary data within several months, wrote a short report, and submitted it in November, 1969, to J. Am. Chem. Soc. as a Communication. It was smoothly accepted, and in the end of that year, a 3-page proof-prints came in. Attached there was a message, saying that a similar study from another laboratory was presently in press and that the corrected proof should be returned as quickly as possible. Thus, we sent it back immediately, and kept watching the journal. A Communication by Shapiro and coworkers appeared in the January 28th issue of the journal and ours in the next issue February 11, 1970.2),3) Both had a diagram of the reaction scheme as shown here in Fig. 5. Their submission was done in October, a month earlier than ours. We were lucky that the journal did not turn down our paper. It was in fact quite fair to publish these almost identical contents from two independent groups in a journal virtually simultaneously.

Bottom Line: At the same time, Shapiro and his coworkers in New York University found the same reaction independently.We also reported that 5-methylcytosine was deaminated by bisulfite only very slowly.The author's recent work that has resulted in an improvement of the procedure of analysis by use of a newly devised high concentration bisulfite solution is also described.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Pharmaceutical Sciences, Okayama University, Okayama, Japan. hayatsuh@cc.okayama-u.ac.jp

ABSTRACT
Methylation at position 5 of cytosine in DNA is being intensively studied in many areas of biological sciences, as the methylation is intimately associated with the control of gene functions. The principal analytical method for determining the sites of 5-methylcytosine in genome at the sequence level involves bisulfite modification of DNA. The utility of this chemical treatment is based on the property of bisulfite to selectively deaminate cytosine residues. The bisulfite-mediated cytosine deamination was discovered in 1970 by us in the University of Tokyo. At the same time, Shapiro and his coworkers in New York University found the same reaction independently. We also reported that 5-methylcytosine was deaminated by bisulfite only very slowly. These findings were later utilized by a group of Australian scientists to devise a means to analyze 5-methylcytosine in DNA; thus, a method called 'bisulfite genomic sequencing' was invented by these researchers in 1992. This review describes the author's reflection of the discovery of bisulfite reactions with pyrimidine bases. The author's recent work that has resulted in an improvement of the procedure of analysis by use of a newly devised high concentration bisulfite solution is also described.

Show MeSH