en
Scientific article
English

Maturation of the head of bacteriophage T4. I. DNA packaging events

Published inJournal of Molecular Biology, vol. 80, no. 4, p. 575-599
Publication date1973-11-15
Abstract

Pulse-chase experiments in wild-type and mutant phage-infected cells provide evidence that the following particles called prohead I, II and III are successive precursors to the mature heads. The prohead I particles contain predominantly the precursor protein P23 and possibly P22 (mol. wt 31,000) and IP III (mol. wt 24,000) and have an s value of about 400 S. Concomitantly with the cleavage of most of P23 (mol. wt 55,000) to P23∗ (mol. wt 45,000), they are rapidly converted into prohead II particles which sediment with about 350 S. The prohead II particles contain, in addition to P23∗, the major constituents of the viral shella—a core consisting of proteins P22 and IP III. In cell lysates, prohead I and prohead II particles contain no DNA in a DNase-resistant form and are not bound to the replicative DNA. We cannot, however, positively rule out the possibility that these particles may have contained some DNA while in the cells.

The prohead II particles are in turn converted into particles which sediment with about 550 S after DNase treatment (prohead III). During this conversion about 50% of normal DNA complement becomes packaged in a DNase-resistant form, and roughly 50% of the core proteins P22 and IP III are cleaved. In lysates the prohead III particles are attached to the replicative DNA. The prohead III particle appears to be the immediate precursor of the full mature head (1100 S). Cleavage of protein P22 to small polypeptides and conversion of IP III IP III∗ are completed at this time. No precursor proteins are found in the full heads. Studies with various mutant phage showed that the prohead II to III conversion is blocked by mutations in genes 16 and 17 and that the conversion of the prohead III particles to the mature heads is blocked by mutations in gene 49. Cleavage of the head proteins, however, occurs normally in these mutant-infected cells. We conclude that the cleavage of the major component of the viral shell, P23, into P23∗ precedes the DNA packaging event, whereas cleavage of the core proteins P22 and IP III appears to be intimately linked to the DNA packaging event. Models relating the cleavage processes to DNA encapsulation are discussed.

eng
Keywords
  • Carbon Radioisotopes
  • Centrifugation, Density Gradient
  • Coliphages / growth & development
  • DNA Replication
  • DNA, Viral
  • Electrophoresis, Disc
  • Escherichia coli
  • Glutaral
  • Glycols
  • Kinetics
  • Leucine / metabolism
  • Methionine / metabolism
  • Molecular Weight
  • Mutation
  • Sulfur Radioisotopes
  • Thymidine / metabolism
  • Tritium
  • Viral Proteins
Affiliation Not a UNIGE publication
Citation (ISO format)
LAEMMLI, Ulrich Karl, FAVRE, M. Maturation of the head of bacteriophage T4. I. DNA packaging events. In: Journal of Molecular Biology, 1973, vol. 80, n° 4, p. 575–599. doi: 10.1016/0022-2836(73)90198-8
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Article (Published version)
accessLevelRestricted
Identifiers
ISSN of the journal0022-2836
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