Biomedical subjects

B Julien

Publications and source records attributed to B Julien.

8 recordsLinked to original sources

PubMed1995

Purification and characterization of the bacteriophage P4 delta protein.

The bacteriophage P4 delta protein is a transcriptional activator of the late genes of P4 as well as the late genes of its helpers, such as bacteriophage P2. delta was purified, using a variation of the MalE fusion system. With this method we purified two forms of delta: a fusion of MalE and delta and a unfused form. The fusion by itself is not active in vivo or in vitro, but the mixture of the fusion and the unfused delta is active in both. Using nitrocellulose filtration and gel mobility shift assays, we show that delta binds DNA, and using DNase I footprinting, we show that delta binds to sequences centered at approximately -55 in the two late promoters of P4 as well as the four late promoters of its helper P2. In addition, the P4 sid promoter contains a second delta binding site centered at -18.

Amino Acid Sequence↗

PubMed1994

Molecular cloning and characterization of bacteriophage P2 genes R and S involved in tail completion.

The sequences of two previously known tail genes, R and S, of the temperate bacteriophage P2 and the sequence of an additional open reading frame (orf-30) located between S and V, were determined. Amber mutations mapping within R and S, Ram3, Ram42, Ram23, Sam75, and Sam89 were sequenced and found to be within their corresponding open reading frames. We constructed overproducing plasmids for R and S and identified these proteins by SDS-PAGE of whole-cell lysates and Coomassie blue staining. The predicted molecular masses of proteins R and S were M(r) 17,400 and 17,300, respectively, although both polypeptides migrated more slowly during gel electrophoresis than would be expected from the sequence data. orf-30 occupies the strand opposite from RS and V and is preceded by several weak potential sigma 70-RNA polymerase promoters, some of which overlap with the V promoter. A construct that had the putative orf-30 promoter region upstream of the lacZ gene produced low levels of beta-galactosidase activity in vivo. Expression from the orf-30 promoter was not stimulated by the phage P4 transcriptional activator protein, delta, which acts at all the known P2 and P4 late promoters. Insertion mutagenesis showed that orf-30 was not an essential gene for P2 growth in Escherichia coli. None of the gene or protein sequences exhibited extensive homology to sequences in the nucleic acid and protein databases. However, the R protein contains a small region homologous to one in the phage T4 tail protein gp15, which is required for T4 tails to bind heads. We propose that R and S are tail completion proteins that are essential for stable head joining.

Amino Acid Sequence↗

PubMed1974