Localization of Polyribosomes Containing Alkaline Phosphatase Nascent Polypeptides on Membranes of Escherichia Coli

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1974 Jan 1

PMID 4587609

Citations 24

Authors

R Cancedda

M J SCHLESINGER

Affiliations

Soon will be listed here.

Abstract

A procedure has been developed for extracting membranes from bacterial cells under conditions that keep a large fraction of bacterial polyribosomes intact. Freeze-thawing spheroplasts in the presence of deoxyribonuclease, followed by differential centrifugation, permits a separation of free and membrane-associated polyribosomes. The latter fraction contains as much as 40% of cell ribosomal ribonucleic acid (RNA) and 55% of cell messenger RNA (mRNA). Nascent polypeptides were divided almost equally between the two fractions, but 70 to 80% of alkaline phosphatase nascent chains, detected both chemically and immunologically, were derived from polyribosomes associated with the bacterial membrane. Analysis of the fractions for mRNA specific for the lac and trp operons by RNA-deoxyribonucleic acid hydridization showed somewhat larger amounts on membrane than on free polyribosomes, but enrichment for nascent alkaline phosphatase (a secreted protein) on membranes was consistently greater, suggesting that polyribosomes making secreted proteins are more tightly bound to membranes. Electron micrographs of the membrane preparations show relatively intact membranes with clusters of polyribosomes on their inner surfaces.

Citing Articles

The Sec System: Protein Export in .

Crane J, Randall L EcoSal Plus. 2017; 7(2).

PMID: 29165233 PMC: 5807066. DOI: 10.1128/ecosalplus.ESP-0002-2017.

ExpA: a conditional mutation affecting the expression of a group of exported proteins in Escherichia coli K-12.

Dassa E, BOQUET P Mol Gen Genet. 1981; 181(2):192-200.

PMID: 7024736 DOI: 10.1007/BF00268426.

Mechanisms of protein localization.

Silhavy T, Benson S, Emr S Microbiol Rev. 1983; 47(3):313-44.

PMID: 6355805 PMC: 281579. DOI: 10.1128/mr.47.3.313-344.1983.

Pleiotropic effects of mutations involved in the regulation of Escherichia coli K-12 alkaline phosphatase.

Morris H, SCHLESINGER M, Bracha M, Yagil E J Bacteriol. 1974; 119(2):583-92.

PMID: 4212247 PMC: 245645. DOI: 10.1128/jb.119.2.583-592.1974.

Both linked and unlinked mutations can alter the intracellular site of synthesis of exported proteins of Escherichia coli.

Rasmussen B, Bassford Jr P J Bacteriol. 1985; 161(1):258-64.

PMID: 3881389 PMC: 214865. DOI: 10.1128/jb.161.1.258-264.1985.

References

Sherr C, UHR J . Immunoglobulin synthesis and secretion. V. Incorporation of leucine and glucosamine into immunoglobulin on free and bound polyribosomes. Proc Natl Acad Sci U S A. 1970; 66(4):1183-9. PMC: 335804. DOI: 10.1073/pnas.66.4.1183. View

Kennell D, Simmons C . Synthesis and decay of messenger ribonucleic acid from the lactose operon of Escherichia coli during amino-acid starvation. J Mol Biol. 1972; 70(3):451-64. DOI: 10.1016/0022-2836(72)90552-9. View

Applebury M, Coleman J . Escherichia coli alkaline phosphatase. Metal binding, protein conformation, and quaternary structure. J Biol Chem. 1969; 244(2):308-18. View

SCHLESINGER M . Activation of a mutationally altered form of the Escherichia coli alkaline phosphatase by zinc. J Biol Chem. 1966; 241(13):3181-8. View

Halford S, Lennette D, Kelley P, SCHLESINGER M . A mutationally altered alkaline phosphatase from Escherichia coli. I. Formation of an active enzyme in vitro and phenotypic suppression in vivo. J Biol Chem. 1972; 247(7):2087-94. View

AMES G . Lipids of Salmonella typhimurium and Escherichia coli: structure and metabolism. J Bacteriol. 1968; 95(3):833-43. PMC: 252100. DOI: 10.1128/jb.95.3.833-843.1968. View

Redman C, Siekevitz P, Palade G . Synthesis and transfer of amylase in pigeon pancreatic micromosomes. J Biol Chem. 1966; 241(5):1150-8. View

Redman C . Biosynthesis of serum proteins and ferritin by free and attached ribosomes of rat liver. J Biol Chem. 1969; 244(16):4308-15. View

Glew R, Heath E . Studies on the extracellular alkaline phosphatase of Micrococcus sodonensis. II. Factors affecting secretion. J Biol Chem. 1971; 246(6):1566-74. View

10.

Milstein C, Brownlee G, Harrison T, Mathews M . A possible precursor of immunoglobulin light chains. Nat New Biol. 1972; 239(91):117-20. DOI: 10.1038/newbio239117a0. View

11.

SCHLESINGER M . Formation of a defective alkaline phosphatase subunit by a mutant of Escherichia coli. J Biol Chem. 1967; 242(7):1604-11. View

12.

HEPPEL L . Selective release of enzymes from bacteria. Science. 1967; 156(3781):1451-5. DOI: 10.1126/science.156.3781.1451. View

13.

Sabatini D, Tashiro Y, Palade G . On the attachment of ribosomes to microsomal membranes. J Mol Biol. 1966; 19(2):503-24. DOI: 10.1016/s0022-2836(66)80019-0. View

14.

Patterson D, Weinstein M, Nixon R, Gillespie D . Interaction of ribosomes and the cell envelope of Escherichia coli mediated by lysozyme. J Bacteriol. 1970; 101(2):584-91. PMC: 284944. DOI: 10.1128/jb.101.2.584-591.1970. View

15.

Coffman R, Norris T, Koch A . Chain elongation rate of messenger and polypeptides in slowly growing Escherichia coli. J Mol Biol. 1971; 60(1):1-19. DOI: 10.1016/0022-2836(71)90442-6. View

16.

Schlesinger S, SCHLESINGER M . The effect of amino acid analogues on alkaline phosphatase formation in Escherichia coli K-12. I. Substitution of triazolealanine for histidine. J Biol Chem. 1967; 242(14):3369-72. View

17.

Vassart G . Specific synthesis of thyroglobulin on membrane bound thyroid ribosomes. FEBS Lett. 1972; 22(1):53-56. DOI: 10.1016/0014-5793(72)80217-5. View

18.

Morris H, SCHLESINGER M . Effects of proline analogues on the formation of alkaline phosphatase in Escherichia coli. J Bacteriol. 1972; 111(1):203-10. PMC: 251259. DOI: 10.1128/jb.111.1.203-210.1972. View

19.

Kennell D, Bicknell I . Decay of messenger ribonucleic acid from the lactose operon of Escherichia coli as a function of growth temperature. J Mol Biol. 1973; 74(1):21-31. DOI: 10.1016/0022-2836(73)90351-3. View

20.

Schlessinger D, MARCHESTI V, Kwan B . BINDING OF RIBOSOMES TO CYTOPLASMIC RETICULUM OF BACILLUS MEGATERIUM. J Bacteriol. 1965; 90:456-66. PMC: 315667. DOI: 10.1128/jb.90.2.456-466.1965. View