Assembly of the Bacteriophage T4 Replication Machine Requires the Acidic Carboxy Terminus of Gene 32 Protein
Identifieur interne : 001313 ( Istex/Curation ); précédent : 001312; suivant : 001314Assembly of the Bacteriophage T4 Replication Machine Requires the Acidic Carboxy Terminus of Gene 32 Protein
Auteurs : J. Michael Hurley [États-Unis] ; Stephen A. Chervitz [États-Unis] ; Thale C. Jarvis [États-Unis] ; Britta S. Singer [États-Unis] ; Larry Gold [États-Unis]Source :
- Journal of Molecular Biology [ 0022-2836 ] ; 1993.
English descriptors
Abstract
Abstract: The acidic carboxy-terminal 89-amino acid fragment of bacteriophage T4 gene 32 protein was expressed in Escherichia coli to high levels from an inducible plasmid construct. Infection of induced cells by wild-type T4 phage results in impaired phage DNA synthesis. The time at which DNA synthesis begins and the diminution in DNA synthesis rates correlate with the amount of carboxy-terminal peptide that accumulates intracellularly prior to infection. Correspondingly, when induced cells are induced cells are infected with viable phage containing a small deletion near the carboxy-terminus of 32 protein (ΔPR201), the inhibition of phage DNA synthesis was much more severe. The mutant 32 protein competes less well against overproduced wild-type acid peptide than does wild-type 32 protein. The purified acid peptide, when used as the attached ligand for affinity chromatography, binds several T4 proteins from phage-infected cells, including 43 protein (T4 DNA polymerase), Dda protein (a DNA helicase), and UvsX protein (a Rec-like recombination protein). Furthermore, at 50- to 100-fold molar excess of acid peptide over intact 32 protein, phage DNA synthesis was specifically inhibited at the initiation step in an in vitro 5-protein DNA replication experiment. We propose that one or more phage replication proteins are titrated as non-productive protein-protein complexes at a site away from the DNA template. This implies that the carboxy-terminal domain of 32 protein is involved in an obligate step of replication machine assembly when the protein is properly attached to ssDNA in the vicinity of a primer-template junction. The assembly defect we observe is strikingly similar to the repression, or "squelching", of the activity of certain eukaryotic transcriptional activators.
Url:
DOI: 10.1006/jmbi.1993.1042
Links toward previous steps (curation, corpus...)
- to stream Istex, to step Corpus: Pour aller vers cette notice dans l'étape Curation :001313
Links to Exploration step
ISTEX:922F3B7724C1197B6A2A2DAA27CCA1E3871ACF75Le document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Assembly of the Bacteriophage T4 Replication Machine Requires the Acidic Carboxy Terminus of Gene 32 Protein</title>
<author><name sortKey="Hurley, J Michael" sort="Hurley, J Michael" uniqKey="Hurley J" first="J. Michael" last="Hurley">J. Michael Hurley</name>
<affiliation wicri:level="1"><mods:affiliation>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309, U.S.A.</mods:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Chervitz, Stephen A" sort="Chervitz, Stephen A" uniqKey="Chervitz S" first="Stephen A." last="Chervitz">Stephen A. Chervitz</name>
<affiliation wicri:level="1"><mods:affiliation>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309, U.S.A.</mods:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Jarvis, Thale C" sort="Jarvis, Thale C" uniqKey="Jarvis T" first="Thale C." last="Jarvis">Thale C. Jarvis</name>
<affiliation wicri:level="1"><mods:affiliation>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309, U.S.A.</mods:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Singer, Britta S" sort="Singer, Britta S" uniqKey="Singer B" first="Britta S." last="Singer">Britta S. Singer</name>
<affiliation wicri:level="1"><mods:affiliation>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309, U.S.A.</mods:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Gold, Larry" sort="Gold, Larry" uniqKey="Gold L" first="Larry" last="Gold">Larry Gold</name>
<affiliation wicri:level="1"><mods:affiliation>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309, U.S.A.</mods:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309</wicri:regionArea>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">ISTEX</idno>
<idno type="RBID">ISTEX:922F3B7724C1197B6A2A2DAA27CCA1E3871ACF75</idno>
<date when="1993" year="1993">1993</date>
<idno type="doi">10.1006/jmbi.1993.1042</idno>
<idno type="url">https://api.istex.fr/ark:/67375/6H6-XMB1GXVC-T/fulltext.pdf</idno>
<idno type="wicri:Area/Istex/Corpus">001313</idno>
<idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">001313</idno>
<idno type="wicri:Area/Istex/Curation">001313</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Assembly of the Bacteriophage T4 Replication Machine Requires the Acidic Carboxy Terminus of Gene 32 Protein</title>
<author><name sortKey="Hurley, J Michael" sort="Hurley, J Michael" uniqKey="Hurley J" first="J. Michael" last="Hurley">J. Michael Hurley</name>
<affiliation wicri:level="1"><mods:affiliation>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309, U.S.A.</mods:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Chervitz, Stephen A" sort="Chervitz, Stephen A" uniqKey="Chervitz S" first="Stephen A." last="Chervitz">Stephen A. Chervitz</name>
<affiliation wicri:level="1"><mods:affiliation>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309, U.S.A.</mods:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Jarvis, Thale C" sort="Jarvis, Thale C" uniqKey="Jarvis T" first="Thale C." last="Jarvis">Thale C. Jarvis</name>
<affiliation wicri:level="1"><mods:affiliation>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309, U.S.A.</mods:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Singer, Britta S" sort="Singer, Britta S" uniqKey="Singer B" first="Britta S." last="Singer">Britta S. Singer</name>
<affiliation wicri:level="1"><mods:affiliation>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309, U.S.A.</mods:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309</wicri:regionArea>
</affiliation>
</author>
<author><name sortKey="Gold, Larry" sort="Gold, Larry" uniqKey="Gold L" first="Larry" last="Gold">Larry Gold</name>
<affiliation wicri:level="1"><mods:affiliation>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309, U.S.A.</mods:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Molecular, Cellular and Developmental Biology University of Colorado, Boulder, CO 80309</wicri:regionArea>
</affiliation>
</author>
</analytic>
<monogr></monogr>
<series><title level="j">Journal of Molecular Biology</title>
<title level="j" type="abbrev">YJMBI</title>
<idno type="ISSN">0022-2836</idno>
<imprint><publisher>ELSEVIER</publisher>
<date type="published" when="1993">1993</date>
<biblScope unit="volume">229</biblScope>
<biblScope unit="issue">2</biblScope>
<biblScope unit="page" from="398">398</biblScope>
<biblScope unit="page" to="418">418</biblScope>
</imprint>
<idno type="ISSN">0022-2836</idno>
</series>
</biblStruct>
</sourceDesc>
<seriesStmt><idno type="ISSN">0022-2836</idno>
</seriesStmt>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>DNA replication</term>
<term>phage T4</term>
<term>protein-protein interactions</term>
<term>squelching</term>
<term>ssDNA binding protein</term>
</keywords>
</textClass>
<langUsage><language ident="en">en</language>
</langUsage>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">Abstract: The acidic carboxy-terminal 89-amino acid fragment of bacteriophage T4 gene 32 protein was expressed in Escherichia coli to high levels from an inducible plasmid construct. Infection of induced cells by wild-type T4 phage results in impaired phage DNA synthesis. The time at which DNA synthesis begins and the diminution in DNA synthesis rates correlate with the amount of carboxy-terminal peptide that accumulates intracellularly prior to infection. Correspondingly, when induced cells are induced cells are infected with viable phage containing a small deletion near the carboxy-terminus of 32 protein (ΔPR201), the inhibition of phage DNA synthesis was much more severe. The mutant 32 protein competes less well against overproduced wild-type acid peptide than does wild-type 32 protein. The purified acid peptide, when used as the attached ligand for affinity chromatography, binds several T4 proteins from phage-infected cells, including 43 protein (T4 DNA polymerase), Dda protein (a DNA helicase), and UvsX protein (a Rec-like recombination protein). Furthermore, at 50- to 100-fold molar excess of acid peptide over intact 32 protein, phage DNA synthesis was specifically inhibited at the initiation step in an in vitro 5-protein DNA replication experiment. We propose that one or more phage replication proteins are titrated as non-productive protein-protein complexes at a site away from the DNA template. This implies that the carboxy-terminal domain of 32 protein is involved in an obligate step of replication machine assembly when the protein is properly attached to ssDNA in the vicinity of a primer-template junction. The assembly defect we observe is strikingly similar to the repression, or "squelching", of the activity of certain eukaryotic transcriptional activators.</div>
</front>
</TEI>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Istex/Curation
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 001313 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Istex/Curation/biblio.hfd -nk 001313 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Sante |area= MersV1 |flux= Istex |étape= Curation |type= RBID |clé= ISTEX:922F3B7724C1197B6A2A2DAA27CCA1E3871ACF75 |texte= Assembly of the Bacteriophage T4 Replication Machine Requires the Acidic Carboxy Terminus of Gene 32 Protein }}
This area was generated with Dilib version V0.6.33. |