Abstract
Transcription from the late promoters of bacteriophage P2 and its satellite phage P4 is activated by a unique class of small, zinc-binding proteins. Using plasmid expression systems, we compared activators from two P2-like (helper) phages with those encoded by two satellite phages. The helper phage activators have more activity on the P4 phage sid promoter. In contrast, the satellite phage activators function better on the four late P2 promoters and on the P4 late leftward promoter. We purified one activator encoded by a P2-like phage and an activator from a satellite phage and determined their binding sites within the P2 and P4 late promoters. Differences in activity levels correlate with binding specificities; promoters that function best with the satellite phage activators have only one activator binding site centered at -55, while the P4 sid promoter, which has more activity with helper phage activators, has a second binding site centered at -18. Surprisingly, DNase I footprinting revealed only very minor differences in promoter binding by the two activators reported here and the P4 activator reported previously. Thus, the differences in transcriptional activity are probably due to interactions between the activators and RNA polymerase, rather than interactions between the activators and DNA.
Full Text
The Full Text of this article is available as a PDF (915.2 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
- Artz S. W., Broach J. R. Histidine regulation in Salmonella typhimurium: an activator attenuator model of gene regulation. Proc Natl Acad Sci U S A. 1975 Sep;72(9):3453–3457. doi: 10.1073/pnas.72.9.3453. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ayers D. J., Sunshine M. G., Six E. W., Christie G. E. Mutations affecting two adjacent amino acid residues in the alpha subunit of RNA polymerase block transcriptional activation by the bacteriophage P2 Ogr protein. J Bacteriol. 1994 Dec;176(24):7430–7438. doi: 10.1128/jb.176.24.7430-7438.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Birkeland N. K., Christie G. E., Lindqvist B. H. Directed mutagenesis of the bacteriophage P2 ogr gene defines an essential function. Gene. 1988 Dec 20;73(2):327–335. doi: 10.1016/0378-1119(88)90497-0. [DOI] [PubMed] [Google Scholar]
- Birkeland N. K., Lindquist B. H. Coliphage P2 late control gene ogr. DNA sequence and product identification. J Mol Biol. 1986 Apr 5;188(3):487–490. doi: 10.1016/0022-2836(86)90170-1. [DOI] [PubMed] [Google Scholar]
- Birkeland N. K., Lindqvist B. H., Christie G. E. Control of bacteriophage P2 gene expression: analysis of transcription of the ogr gene. J Bacteriol. 1991 Nov;173(21):6927–6934. doi: 10.1128/jb.173.21.6927-6934.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bullas L. R., Mostaghimi A. R., Arensdorf J. J., Rajadas P. T., Zuccarelli A. J. Salmonella phage PSP3, another member of the P2-like phage group. Virology. 1991 Dec;185(2):918–921. doi: 10.1016/0042-6822(91)90573-t. [DOI] [PubMed] [Google Scholar]
- Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Christie G. E., Calendar R. Bacteriophage P2 late promoters. II. Comparison of the four late promoter sequences. J Mol Biol. 1985 Feb 5;181(3):373–382. doi: 10.1016/0022-2836(85)90226-8. [DOI] [PubMed] [Google Scholar]
- Christie G. E., Calendar R. Bacteriophage P2 late promoters. Transcription initiation sites for two late mRNAs. J Mol Biol. 1983 Jul 15;167(4):773–790. doi: 10.1016/s0022-2836(83)80110-7. [DOI] [PubMed] [Google Scholar]
- Christie G. E., Haggård-Ljungquist E., Feiwell R., Calendar R. Regulation of bacteriophage P2 late-gene expression: the ogr gene. Proc Natl Acad Sci U S A. 1986 May;83(10):3238–3242. doi: 10.1073/pnas.83.10.3238. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Collado-Vides J., Magasanik B., Gralla J. D. Control site location and transcriptional regulation in Escherichia coli. Microbiol Rev. 1991 Sep;55(3):371–394. doi: 10.1128/mr.55.3.371-394.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dale E. C., Christie G. E., Calendar R. Organization and expression of the satellite bacteriophage P4 late gene cluster. J Mol Biol. 1986 Dec 20;192(4):793–803. doi: 10.1016/0022-2836(86)90029-x. [DOI] [PubMed] [Google Scholar]
- Dehó G., Zangrossi S., Ghisotti D., Sironi G. Alternative promoters in the development of bacteriophage plasmid P4. J Virol. 1988 May;62(5):1697–1704. doi: 10.1128/jvi.62.5.1697-1704.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dibbens J. A., Gregory S. L., Egan J. B. Control of gene expression in the temperate coliphage 186. X. The cI repressor directly represses transcription of the late control gene B. Mol Microbiol. 1992 Sep;6(18):2643–2650. doi: 10.1111/j.1365-2958.1992.tb01441.x. [DOI] [PubMed] [Google Scholar]
- Dunbar B. S., Kimura H., Timmons T. M. Protein analysis using high-resolution two-dimensional polyacrylamide gel electrophoresis. Methods Enzymol. 1990;182:441–459. doi: 10.1016/0076-6879(90)82036-2. [DOI] [PubMed] [Google Scholar]
- Ghisotti D., Finkel S., Halling C., Dehò G., Sironi G., Calendar R. Nonessential region of bacteriophage P4: DNA sequence, transcription, gene products, and functions. J Virol. 1990 Jan;64(1):24–36. doi: 10.1128/jvi.64.1.24-36.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Grambow N. J., Birkeland N. K., Anders D. L., Christie G. E. Deletion analysis of a bacteriophage P2 late promoter. Gene. 1990 Oct 30;95(1):9–15. doi: 10.1016/0378-1119(90)90407-i. [DOI] [PubMed] [Google Scholar]
- Griffin T. J., 4th, Kolodner R. D. Purification and preliminary characterization of the Escherichia coli K-12 recF protein. J Bacteriol. 1990 Nov;172(11):6291–6299. doi: 10.1128/jb.172.11.6291-6299.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Halling C., Sunshine M. G., Lane K. B., Six E. W., Calendar R. A mutation of the transactivation gene of satellite bacteriophage P4 that suppresses the rpoA109 mutation of Escherichia coli. J Bacteriol. 1990 Jul;172(7):3541–3548. doi: 10.1128/jb.172.7.3541-3548.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ishihama A. Protein-protein communication within the transcription apparatus. J Bacteriol. 1993 May;175(9):2483–2489. doi: 10.1128/jb.175.9.2483-2489.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jin S., Chen Y., Christie G. E., Benedik M. J. Regulation of the Serratia marcescens extracellular nuclease: positive control by a homolog of P2 Ogr encoded by a cryptic prophage. J Mol Biol. 1996 Feb 23;256(2):264–278. doi: 10.1006/jmbi.1996.0084. [DOI] [PubMed] [Google Scholar]
- Kahn M. L., Ziermann R., Dehò G., Ow D. W., Sunshine M. G., Calendar R. Bacteriophage P2 and P4. Methods Enzymol. 1991;204:264–280. doi: 10.1016/0076-6879(91)04013-e. [DOI] [PubMed] [Google Scholar]
- Kalionis B., Dodd I. B., Egan J. B. Control of gene expression in the P2-related template coliphages. III. DNA sequence of the major control region of phage 186. J Mol Biol. 1986 Sep 20;191(2):199–209. doi: 10.1016/0022-2836(86)90257-3. [DOI] [PubMed] [Google Scholar]
- Kalionis B., Pritchard M., Egan J. B. Control of gene expression in the P2-related temperate coliphages. IV. Concerning the late control gene and control of its transcription. J Mol Biol. 1986 Sep 20;191(2):211–220. doi: 10.1016/0022-2836(86)90258-5. [DOI] [PubMed] [Google Scholar]
- Keener J., Dale E. C., Kustu S., Calendar R. In vitro transcription from the late promoter of bacteriophage P4. J Bacteriol. 1988 Aug;170(8):3543–3546. doi: 10.1128/jb.170.8.3543-3546.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kumar A., Grimes B., Fujita N., Makino K., Malloch R. A., Hayward R. S., Ishihama A. Role of the sigma 70 subunit of Escherichia coli RNA polymerase in transcription activation. J Mol Biol. 1994 Jan 14;235(2):405–413. doi: 10.1006/jmbi.1994.1001. [DOI] [PubMed] [Google Scholar]
- Lee T. C., Christie G. E. Purification and properties of the bacteriophage P2 ogr gene product. A prokaryotic zinc-binding transcriptional activator. J Biol Chem. 1990 May 5;265(13):7472–7477. [PubMed] [Google Scholar]
- Lessl M., Balzer D., Lurz R., Waters V. L., Guiney D. G., Lanka E. Dissection of IncP conjugative plasmid transfer: definition of the transfer region Tra2 by mobilization of the Tra1 region in trans. J Bacteriol. 1992 Apr;174(8):2493–2500. doi: 10.1128/jb.174.8.2493-2500.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Li M., Moyle H., Susskind M. M. Target of the transcriptional activation function of phage lambda cI protein. Science. 1994 Jan 7;263(5143):75–77. doi: 10.1126/science.8272867. [DOI] [PubMed] [Google Scholar]
- Lindqvist B. H., Six E. W. Replication of bacteriophage P4 DNA in a nonlysogenic host. Virology. 1971 Jan;43(1):1–7. doi: 10.1016/0042-6822(71)90218-2. [DOI] [PubMed] [Google Scholar]
- Lisser S., Margalit H. Compilation of E. coli mRNA promoter sequences. Nucleic Acids Res. 1993 Apr 11;21(7):1507–1516. doi: 10.1093/nar/21.7.1507. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Makino K., Amemura M., Kim S. K., Nakata A., Shinagawa H. Role of the sigma 70 subunit of RNA polymerase in transcriptional activation by activator protein PhoB in Escherichia coli. Genes Dev. 1993 Jan;7(1):149–160. doi: 10.1101/gad.7.1.149. [DOI] [PubMed] [Google Scholar]
- Neidhardt F. C., Bloch P. L., Smith D. F. Culture medium for enterobacteria. J Bacteriol. 1974 Sep;119(3):736–747. doi: 10.1128/jb.119.3.736-747.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pritchard M., Egan J. B. Control of gene expression in P2-related coliphages: the in vitro transcription pattern of coliphage 186. EMBO J. 1985 Dec 16;4(13A):3599–3604. doi: 10.1002/j.1460-2075.1985.tb04123.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sadler J. R., Tecklenburg M., Betz J. L. Plasmids containing many tandem copies of a synthetic lactose operator. Gene. 1980 Feb;8(3):279–300. doi: 10.1016/0378-1119(80)90005-0. [DOI] [PubMed] [Google Scholar]
- Sedmak J. J., Grossberg S. E. A rapid, sensitive, and versatile assay for protein using Coomassie brilliant blue G250. Anal Biochem. 1977 May 1;79(1-2):544–552. doi: 10.1016/0003-2697(77)90428-6. [DOI] [PubMed] [Google Scholar]
- Siegele D. A., Hu J. C., Walter W. A., Gross C. A. Altered promoter recognition by mutant forms of the sigma 70 subunit of Escherichia coli RNA polymerase. J Mol Biol. 1989 Apr 20;206(4):591–603. doi: 10.1016/0022-2836(89)90568-8. [DOI] [PubMed] [Google Scholar]
- Simons R. W., Houman F., Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. doi: 10.1016/0378-1119(87)90095-3. [DOI] [PubMed] [Google Scholar]
- Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
- Sun J., Inouye M., Inouye S. Association of a retroelement with a P4-like cryptic prophage (retronphage phi R73) integrated into the selenocystyl tRNA gene of Escherichia coli. J Bacteriol. 1991 Jul;173(13):4171–4181. doi: 10.1128/jb.173.13.4171-4181.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Van Bokkelen G. B., Dale E. C., Halling C., Calendar R. Mutational analysis of a bacteriophage P4 late promoter. J Bacteriol. 1991 Jan;173(1):37–45. doi: 10.1128/jb.173.1.37-45.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]
- Waldburger C., Gardella T., Wong R., Susskind M. M. Changes in conserved region 2 of Escherichia coli sigma 70 affecting promoter recognition. J Mol Biol. 1990 Sep 20;215(2):267–276. doi: 10.1016/s0022-2836(05)80345-6. [DOI] [PubMed] [Google Scholar]