2024-03-29T14:40:55Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/393082018-10-17T08:28:32Zcom_10261_79com_10261_1col_10261_332
DIGITAL.CSIC
author
Garmendia Mendizábal, Cristina
author
Hermoso, José Miguel
author
Salas, Margarita
funder
National Institutes of Health (US)
funder
Ministerio de Economía y Competitividad (España)
funder
Fundación Ramón Areces
2011-09-06T12:44:33Z
2011-09-06T12:44:33Z
1990-03-30
Gene 88(1): 73-79 (1990)
0378-1119
http://hdl.handle.net/10261/39308
10.1016/0378-1119(90)90061-U
http://dx.doi.org/10.13039/100000002http://dx.doi.org/10.13039/501100003329http://dx.doi.org/10.13039/100008054
By site-directed mutagenesis we have changed into Cys the Ser232 of the φ29 terminal protein (TP) involved in the covalent linkage to dAMP for the initiation of replication. The mutant TP, highly purified, had about 0.7% of the priming activity of the wild-type (wt) protein p3. The linkage between the mutant protein p3 and dAMP was more labile to piperidine treatment than the serine-dAMP linkage in the wt protein p3, suggesting the presence of a different kind of linkage, Cys-dAMP. In the other three mutant TPs, residues Leu220, Ser223 and Ser226 were independently changed into Pro; the purified TP mutants had about 3%, 140% and 1% of the priming activity of the wt p3, respectively. All the mutant TP were able to interact with the φ29 DNA polymerase and with DNA, suggesting that Leu220 and Ser266, in addition to Ser232, form part of a functional domain involved in the process of initiation of DNA replication.
eng
closedAccess
Protein p3-dAMP complex
Site directed mutagenesis
Initiation of DNA replication
DNA polymerase
Functional domain for priming activity in the phage φ29 terminal protein
artículo
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