Abstract
The Escherichia coli dnaK gene product, originally defined by mutations that blocked λ phage DNA replication, is known to be necessary for E. coli viability. We have purified dnaK protein to homogeneity and have demonstrated that it possesses a weak DNA-independent ATPase activity, which results in the production of ADP and P(i). The proof that this ATPase activity is encoded by the dnaK+ gene relies primarily on the fact that the dnaK756 mutation results in the production of an ATPase activity with altered physical properties. The dnaK protein is phosphorylated in vitro and in vivo, probably as a result of an autophosphorylation reaction. The λ O and P replication proteins were shown to interact in vitro with the dnaK protein. The ATPase activity of the dnaK protein was inhibited by purified λ P protein and stimulated by purified λ O protein. Moreover, the dnaK protein participates in the initiation of DNA synthesis in an in vitro DNA replication system that is dependent on the O and P proteins. Anti-dnaK protein immunoglobulin specifically inhibited DNA synthesis in this system.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 6431-6435 |
| Number of pages | 5 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 80 |
| Issue number | 21 I |
| State | Published - 1983 |
| Externally published | Yes |
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ASJC Scopus subject areas
- General
- Genetics
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The dnaK protein of Escherichia coli possesses an ATPase and autophosphorylating activity and is essential in an in vitro DNA replication system. / Zylicz, M.; LeBowitz, J. H.; McMacken, Roger; Georgopoulos, C.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 80, No. 21 I, 1983, p. 6431-6435.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - The dnaK protein of Escherichia coli possesses an ATPase and autophosphorylating activity and is essential in an in vitro DNA replication system
AU - Zylicz, M.
AU - LeBowitz, J. H.
AU - McMacken, Roger
AU - Georgopoulos, C.
PY - 1983
Y1 - 1983
N2 - The Escherichia coli dnaK gene product, originally defined by mutations that blocked λ phage DNA replication, is known to be necessary for E. coli viability. We have purified dnaK protein to homogeneity and have demonstrated that it possesses a weak DNA-independent ATPase activity, which results in the production of ADP and P(i). The proof that this ATPase activity is encoded by the dnaK+ gene relies primarily on the fact that the dnaK756 mutation results in the production of an ATPase activity with altered physical properties. The dnaK protein is phosphorylated in vitro and in vivo, probably as a result of an autophosphorylation reaction. The λ O and P replication proteins were shown to interact in vitro with the dnaK protein. The ATPase activity of the dnaK protein was inhibited by purified λ P protein and stimulated by purified λ O protein. Moreover, the dnaK protein participates in the initiation of DNA synthesis in an in vitro DNA replication system that is dependent on the O and P proteins. Anti-dnaK protein immunoglobulin specifically inhibited DNA synthesis in this system.
AB - The Escherichia coli dnaK gene product, originally defined by mutations that blocked λ phage DNA replication, is known to be necessary for E. coli viability. We have purified dnaK protein to homogeneity and have demonstrated that it possesses a weak DNA-independent ATPase activity, which results in the production of ADP and P(i). The proof that this ATPase activity is encoded by the dnaK+ gene relies primarily on the fact that the dnaK756 mutation results in the production of an ATPase activity with altered physical properties. The dnaK protein is phosphorylated in vitro and in vivo, probably as a result of an autophosphorylation reaction. The λ O and P replication proteins were shown to interact in vitro with the dnaK protein. The ATPase activity of the dnaK protein was inhibited by purified λ P protein and stimulated by purified λ O protein. Moreover, the dnaK protein participates in the initiation of DNA synthesis in an in vitro DNA replication system that is dependent on the O and P proteins. Anti-dnaK protein immunoglobulin specifically inhibited DNA synthesis in this system.
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UR - http://www.scopus.com/inward/citedby.url?scp=0043104253&partnerID=8YFLogxK
M3 - Article
C2 - 6314326
AN - SCOPUS:0043104253
VL - 80
SP - 6431
EP - 6435
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 21 I
ER -