Formation of a covalent complex between the terminal protein of pneumococcal bacteriophage Cp-1 and 5'-dAMP

Journal Article · Tue Apr 01 00:00:00 EST 1986 · J. Virol.; (United States)

OSTI ID:5207452

Garcia, P; Hermoso, J M; Garcia, J A; Garcia, E; Lopez, R; Salas, M

Incubation of extracts of Cp-1-infected Streptococcus pneumoniae with (..cap alpha..-/sup 32/P)dATP produced a labeled protein with the electrophoretic mobility of the Cp-1 terminal protein. The reaction product was resistant to treatment with micrococcal nuclease and sensitive to treatment with proteinase K. Incubation of the /sup 32/P-labeled protein with 5 M piperidine for 4 h at 50/sup 0/C released 5'-dAMP, indicating that a covalent complex between the terminal protein and 5'-dAMP was formed in vitro. When the four deoxynucleoside triphosphates were included in the reaction mixture, a labeled complex of slower electrophoretic mobility in sodium dodecyl sulfate-polyacrylamide gels than the terminal protein-dAMP complex was also found, indicating that the Cp-1 terminal protein-dAMP complex can be elongated and, therefore, that it is an initiation complex. Treatment of the /sup 32/P-labeled terminal protein-dAMP complex with 5.8 M HCl at 110/sup 0/C for 2 h yielded phosphothreonine. These results, together with the resistance of the terminal protein-DNA linkage to hydroxylamine, suggest that the Cp-1 terminal protein is covalently linked to the DNA through a phosphoester bond between L-threonine and 5'-dAMP, namely, a O-5'-deoxyadenylyl-L-threonine bond.

• Research Organization: Instituto de Inmunologia y Biologia Microbiana, Madrid, Spain
• OSTI ID: 5207452
• Journal Information: J. Virol.; (United States), Vol. 58:1
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
PROTEINS
BIOCHEMICAL REACTION KINETICS
AMP
ATP
BACTERIOPHAGES
COVALENCE
ELECTROPHORESIS
NUCLEASES
PEPTIDE HYDROLASES
PHOSPHORUS 32
PIPERIDINES
STREPTOCOCCUS
TRACER TECHNIQUES
AMINES
AZINES
BACTERIA
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
DAYS LIVING RADIOISOTOPES
ENZYMES
ESTERASES
HETEROCYCLIC COMPOUNDS
HYDROLASES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LIGHT NUCLEI
MICROORGANISMS
NUCLEI
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
PARASITES
PHOSPHODIESTERASES
PHOSPHORUS ISOTOPES
PYRIDINES
RADIOISOTOPES
REACTION KINETICS
VIRUSES
550201* - Biochemistry- Tracer Techniques