Overproduction of Escherichia Coli Integration Host Factor, a Protein with Nonidentical Subunits

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1987 Sep 1

PMID 3305480

Citations 80

Authors

H A NASH

C A Robertson

E Flamm

R A WEISBERG

H I MILLER

Affiliations

Soon will be listed here.

Abstract

Integration host factor (IHF) is a small, basic protein that is needed for efficient recombination of bacteriophage lambda, as well as for other host and viral functions. We have constructed strains in which the two subunits of IHF, encoded by the himA and hip genes of Escherichia coli, are expressed under the control of the lambda rho L promoter. Separate overexpression of himA and hip led to the production of unstable and insoluble peptides, respectively. In contrast, the overexpression of both genes conjointly led to the accumulation of large amounts of active IHF. Extracts of such cells provided the starting material for a rapid purification procedure that results in milligram quantities of apparently homogeneous IHF.

Citing Articles

Reciprocally rewiring and repositioning the Integration Host Factor (IHF) subunit genes in serovar Typhimurium: impacts on physiology and virulence.

Pozdeev G, Beckett M, Mogre A, Thomson N, Dorman C Microb Genom. 2022; 8(2).

PMID: 35166652 PMC: 8942017. DOI: 10.1099/mgen.0.000768.

Endogenous and Foreign Nucleoid-Associated Proteins of Bacteria: Occurrence, Interactions and Effects on Mobile Genetic Elements and Host's Biology.

Flores-Rios R, Quatrini R, Loyola A Comput Struct Biotechnol J. 2019; 17:746-756.

PMID: 31303979 PMC: 6606824. DOI: 10.1016/j.csbj.2019.06.010.

Three tandem promoters, together with IHF, regulate growth phase dependent expression of the Escherichia coli kps capsule gene cluster.

Jia J, King J, Goldrick M, Aldawood E, Roberts I Sci Rep. 2017; 7(1):17924.

PMID: 29263430 PMC: 5738388. DOI: 10.1038/s41598-017-17891-0.

The interplay between DNA topology and accessory factors in site-specific recombination in bacteria and their bacteriophages.

Dorman C, Bogue M Sci Prog. 2017; 99(4):420-437.

PMID: 28742481 PMC: 10365484. DOI: 10.3184/003685016X14811202974921.

Physical and Functional Characterization of a Viral Genome Maturation Complex.

Yang T, Ortiz D, Yang Q, De Angelis R, Sanyal S, Catalano C Biophys J. 2017; 112(8):1551-1560.

PMID: 28445747 PMC: 5406279. DOI: 10.1016/j.bpj.2017.02.041.

References

Mizuuchi K, Gellert M, NASH H . Involement of supertwisted DNA in integrative recombination of bacteriophage lambda. J Mol Biol. 1978; 121(3):375-92. DOI: 10.1016/0022-2836(78)90370-4. View

Kikuchi Y, NASH H . The bacteriophage lambda int gene product. A filter assay for genetic recombination, purification of int, and specific binding to DNA. J Biol Chem. 1978; 253(20):7149-57. View

MILLER H, Kikuchi A, NASH H, WEISBERG R, Friedman D . Site-specific recombination of bacteriophage lambda: the role of host gene products. Cold Spring Harb Symp Quant Biol. 1979; 43 Pt 2:1121-6. DOI: 10.1101/sqb.1979.043.01.125. View

Birnboim H, DOLY J . A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979; 7(6):1513-23. PMC: 342324. DOI: 10.1093/nar/7.6.1513. View

MILLER H, Friedman D . An E. coli gene product required for lambda site-specific recombination. Cell. 1980; 20(3):711-9. DOI: 10.1016/0092-8674(80)90317-7. View

NASH H, Robertson C . Purification and properties of the Escherichia coli protein factor required for lambda integrative recombination. J Biol Chem. 1981; 256(17):9246-53. View

Hoyt M, Knight D, Das A, MILLER H, Echols H . Control of phage lambda development by stability and synthesis of cII protein: role of the viral cIII and host hflA, himA and himD genes. Cell. 1982; 31(3 Pt 2):565-73. DOI: 10.1016/0092-8674(82)90312-9. View

NASH H . Purification and properties of the bacteriophage lambda Int protein. Methods Enzymol. 1983; 100:210-6. DOI: 10.1016/0076-6879(83)00057-9. View

NASH H . Purification and properties of Int-h, a variant protein involved in site-specific recombination of bacteriophage lambda. J Biol Chem. 1984; 259(20):12724-32. View

10.

Craig N, NASH H . E. coli integration host factor binds to specific sites in DNA. Cell. 1984; 39(3 Pt 2):707-16. DOI: 10.1016/0092-8674(84)90478-1. View

11.

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

12.

MILLER H . Primary structure of the himA gene of Escherichia coli: homology with DNA-binding protein HU and association with the phenylalanyl-tRNA synthetase operon. Cold Spring Harb Symp Quant Biol. 1984; 49:691-8. DOI: 10.1101/sqb.1984.049.01.078. View

13.

Flamm E, WEISBERG R . Primary structure of the hip gene of Escherichia coli and of its product, the beta subunit of integration host factor. J Mol Biol. 1985; 183(2):117-28. DOI: 10.1016/0022-2836(85)90206-2. View

14.

GARDNER J, NASH H . Role of Escherichia coli IHF protein in lambda site-specific recombination. A mutational analysis of binding sites. J Mol Biol. 1986; 191(2):181-9. DOI: 10.1016/0022-2836(86)90255-x. View

15.

Drlica K, Rouviere-Yaniv J . Histonelike proteins of bacteria. Microbiol Rev. 1987; 51(3):301-19. PMC: 373113. DOI: 10.1128/mr.51.3.301-319.1987. View