Analysis of NutR, a Site Required for Transcription Antitermination in Phage Lambda

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1987 Jul 1

PMID 2955408

Citations 18

Authors

M Zuber

T A Patterson

D L Court

Affiliations

Soon will be listed here.

Abstract

Deletions extending from the cro gene into boxA and nutR of the Rho-dependent tR1 terminator of bacteriophage lambda have been generated and cloned between promoters and the galK gene of Escherichia coli on a multicopy plasmid. Terminators placed between the promoters and galK restrict transcription and expression of galK on these plasmids. However, when lambda N protein is provided, and if a functional N interaction site, nutR, is intact, transcription antitermination occurs and galK expression increases. Deletions into the nutR region affect the ability to antiterminate. From the results obtained we conclude that: boxA, a site believed to bind host factors (Nus), is not required for transcription antitermination in this system; the host NusA function is required even in the absence of boxA; nutR is required for N antitermination; translation across the nutR sequence prevents N-dependent antitermination.

Citing Articles

RNA-binding specificity of E. coli NusA.

Prasch S, Jurk M, Washburn R, Gottesman M, Wohrl B, Rosch P Nucleic Acids Res. 2009; 37(14):4736-42.

PMID: 19515940 PMC: 2724277. DOI: 10.1093/nar/gkp452.

Biochemical and structural characterization of the secreted chorismate mutase (Rv1885c) from Mycobacterium tuberculosis H37Rv: an *AroQ enzyme not regulated by the aromatic amino acids.

Kim S, Reddy S, Nelson B, Vasquez G, Davis A, Howard A J Bacteriol. 2006; 188(24):8638-48.

PMID: 17146044 PMC: 1698256. DOI: 10.1128/JB.00441-06.

Crystal structure of the YajQ protein from Haemophilus influenzae reveals a tandem of RNP-like domains.

Teplyakov A, Obmolova G, Bir N, Reddy P, Howard A, Gilliland G J Struct Funct Genomics. 2003; 4(1):1-9.

PMID: 12943362 DOI: 10.1023/a:1024620416876.

Cloning and expression of the gene for a novel protein from Mycobacterium smegmatis with functional similarity to eukaryotic calmodulin.

Reddy P, Prasad C, Reddy P, Reeder D, McKenney K, Jaffe H J Bacteriol. 2003; 185(17):5263-8.

PMID: 12923099 PMC: 180971. DOI: 10.1128/JB.185.17.5263-5268.2003.

Modulation of Escherichia coli adenylyl cyclase activity by catalytic-site mutants of protein IIA(Glc) of the phosphoenolpyruvate:sugar phosphotransferase system.

Reddy P, Kamireddi M J Bacteriol. 1998; 180(3):732-6.

PMID: 9457881 PMC: 106945. DOI: 10.1128/JB.180.3.732-736.1998.

References

Rosenberg M, Court D, Shimatake H, Brady C, WULFF D . The relationship between function and DNA sequence in an intercistronic regulatory region in phage lambda. Nature. 1978; 272(5652):414-23. DOI: 10.1038/272414a0. View

Montanez C, Bueno J, Schmeissner U, Court D, Guarneros G . Mutations of bacteriophage lambda that define independent but overlapping RNA processing and transcription termination sites. J Mol Biol. 1986; 191(1):29-37. DOI: 10.1016/0022-2836(86)90420-1. View

de Crombrugghe B, Mudryj M, DiLauro R, Gottesman M . Specificity of the bacteriophage lambda N gene product (pN): nut sequences are necessary and sufficient for antitermination by pN. Cell. 1979; 18(4):1145-51. DOI: 10.1016/0092-8674(79)90227-7. View

Maxam A, Gilbert W . Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980; 65(1):499-560. DOI: 10.1016/s0076-6879(80)65059-9. View

GOTTESMAN M, Adhya S, Das A . Transcription antitermination by bacteriophage lambda N gene product. J Mol Biol. 1980; 140(1):57-75. DOI: 10.1016/0022-2836(80)90356-3. View

Gold L, Pribnow D, Schneider T, Shinedling S, Singer B, Stormo G . Translational initiation in prokaryotes. Annu Rev Microbiol. 1981; 35:365-403. DOI: 10.1146/annurev.mi.35.100181.002053. View

Drahos D, Galluppi G, Caruthers M, SZYBALSKI W . Synthesis of the nutL DNA segments and analysis of antitermination and termination functions in coliphage lambda. Gene. 1982; 18(3):343-54. DOI: 10.1016/0378-1119(82)90173-1. View

Olson E, Flamm E, Friedman D . Analysis of nutR: a region of phage lambda required for antitermination of transcription. Cell. 1982; 31(1):61-70. DOI: 10.1016/0092-8674(82)90405-6. View

Irani M, Orosz L, Adhya S . A control element within a structural gene: the gal operon of Escherichia coli. Cell. 1983; 32(3):783-8. DOI: 10.1016/0092-8674(83)90064-8. View

10.

Dambly-Chaudiere C, Gottesman M, Debouck C, Adhya S . Regulation of the pR operon of bacteriophage lambda. J Mol Appl Genet. 1983; 2(1):45-56. View

11.

Friedman D, Olson E . Evidence that a nucleotide sequence, "boxA," is involved in the action of the NusA protein. Cell. 1983; 34(1):143-9. DOI: 10.1016/0092-8674(83)90144-7. View

12.

Guo L, Wu R . Exonuclease III: use for DNA sequence analysis and in specific deletions of nucleotides. Methods Enzymol. 1983; 100:60-96. DOI: 10.1016/0076-6879(83)00046-4. View

13.

Warren F, Das A . Formation of termination-resistant transcription complex at phage lambda nut locus: effects of altered translation and a ribosomal mutation. Proc Natl Acad Sci U S A. 1984; 81(12):3612-6. PMC: 345268. DOI: 10.1073/pnas.81.12.3612. View

14.

Das A, Wolska K . Transcription antitermination in vitro by lambda N gene product: requirement for a phage nut site and the products of host nusA, nusB, and nusE genes. Cell. 1984; 38(1):165-73. DOI: 10.1016/0092-8674(84)90537-3. View

15.

Friedman D, Olson E, Georgopoulos C, Tilly K, Herskowitz I, Banuett F . Interactions of bacteriophage and host macromolecules in the growth of bacteriophage lambda. Microbiol Rev. 1984; 48(4):299-325. PMC: 373221. DOI: 10.1128/mr.48.4.299-325.1984. View

16.

Olson E, Tomich C, Friedman D . The nusA recognition site. Alteration in its sequence or position relative to upstream translation interferes with the action of the N antitermination function of phage lambda. J Mol Biol. 1984; 180(4):1053-63. DOI: 10.1016/0022-2836(84)90270-5. View

17.

Peltz S, Brown A, Hasan N, Podhajska A, SZYBALSKI W . Thermosensitivity of a DNA recognition site: activity of a truncated nutL antiterminator of coliphage lambda. Science. 1985; 228(4695):91-3. DOI: 10.1126/science.3156406. View

18.

Grayhack E, Yang X, Lau L, Roberts J . Phage lambda gene Q antiterminator recognizes RNA polymerase near the promoter and accelerates it through a pause site. Cell. 1985; 42(1):259-69. DOI: 10.1016/s0092-8674(85)80121-5. View

19.

Tsugawa A, Kurihara T, Zuber M, Court D, Nakamura Y . E. coli NusA protein binds in vitro to an RNA sequence immediately upstream of the boxA signal of bacteriophage lambda. EMBO J. 1985; 4(9):2337-42. PMC: 554506. DOI: 10.1002/j.1460-2075.1985.tb03935.x. View

20.

Brown A, SZYBALSKI W . Transcriptional antitermination activity of the synthetic nut elements of coliphage lambda. I. Assembly of the nutR recognition site from boxA and nut core elements. Gene. 1986; 42(1):E125-32. DOI: 10.1016/0378-1119(86)90159-9. View