Polyribosomes and DNA-dependent amino acid incorporation in Escherichia coli extracts

Eiserling, Frederick A.; Levin, Joseph; Byrne, Raymond; Karlsson, Ulf; Nirenberg, M. W.; Sjoestrand, F S

doi:10.1016/s0022-2836(64)80073-5

Cited by 17 publications

(5 citation statements)

References 12 publications

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“…This pattern could have arisen by the dehydration of a helically wound polysome such as that inferred from electron micrographs of thinly sectioned cytoplasm from various tissues. Eiserling et al (5) as well as Shelton and Kuff (19) showed that smaller polysomes often formed rosettes or circles, with the smaller ribosomal subunit in the center. This configuration would also maximize the interaction of adjacent ribosomal subunits.…”

Section: Resultsmentioning

confidence: 99%

“…There have been many electron microscopic studies of these structures in tissue sections, but comparatively few studies have been reported on polysomes after they have been isolated from cells (21,19). Studies of protein synthesis are frequently carried out in Escherichia coli but, although electron microscopic studies have been done on ribosomes (6,8) and small polysomes (5), there have been no reports on the morphology of large polysomes of E. coli. In the course of studying the appearance of the inducible enzyme j3-galactosidase on the polysomes of E. coli, gentle methods of cell lysis were developed which made it possible for us to prepare what appeared to be largely undegraded polysomes (11).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

An Electron Microscopic Study of Large Bacterial Polyribosomes

Slayter

Kiho

Hall

et al. 1968

The Journal of Cell Biology

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

An Electron Microscopic Study of Large Bacterial Polyribosomes

Slayter

Kiho

Hall

et al. 1968

The Journal of Cell Biology

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“…3 At UCLA, Bob Romig often took his graduate students across town to Caltech for important seminars where we met Max Delbruck, Bob Edgar, Dick Epstein, Jean Weigle and a seminar visitor from Geneva, Edouard Kellenberger. Professor Sjostrand was interested in ribosomes, and he sent me to NIH to do microscopy of ribosomes bound to mRNA working with Phillip Leder's group in the lab of Marshall Nirenberg.…”

Section: Life In Sciencementioning

confidence: 99%

Life in science

Eiserling

2015

Bacteriophage

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“…The recent development of cell-free expression is very useful for the study of membrane protein folding. Although the method itself has been known for decades ( Eiserling et al, 1964 ), the overall synthesis efficiency was improved considerably by refinement of protocols and reaction designs ( Henrich et al, 2015 ). This system has been successfully applied to studies of co-translational folding due to facile operation and large flexibility ( Pellowe and Booth, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

Monitoring the Progression of Cell-Free Expression of Microbial Rhodopsins by Surface Enhanced IR Spectroscopy: Resolving a Branch Point for Successful/Unsuccessful Folding

Ataka

Baumann

Chen

et al. 2022

Front. Mol. Biosci.

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The translocon-unassisted folding process of transmembrane domains of the microbial rhodopsins sensory rhodopsin I (HsSRI) and II (HsSRII), channelrhodopsin II (CrChR2), and bacteriorhodopsin (HsBR) during cell-free expression has been investigated by Surface-Enhanced Infrared Absorption Spectroscopy (SEIRAS). Up to now, only a limited number of rhodopsins have been expressed and folded into the functional holoprotein in cell free expression systems, while other microbial rhodopsins fail to properly bind the chromophore all-trans retinal as indicated by the missing visible absorption. SEIRAS experiments suggest that all investigated rhodopsins lead to the production of polypeptides, which are co-translationally inserted into a solid-supported lipid bilayer during the first hour after the in-vitro expression is initiated. Secondary structure analysis of the IR spectra revealed that the polypeptides form a comparable amount of α-helical structure during the initial phase of insertion into the lipid bilayer. As the process progressed (>1 h), only HsBR exhibited a further increase and association of α-helices to form a compact tertiary structure, while the helical contents of the other rhodopsins stagnated. This result suggests that the molecular reason for the unsuccessful cell-free expression of the two sensory rhodopsins and of CrChR2 is not due to the translation process, but rather to the folding process during the post-translational period. Taking our previous observation into account that HsBR fails to form a tertiary structure in the absence of its retinal, we infer that the chromophore retinal is an integral component of the compaction of the polypeptide into its tertiary structure and the formation of a fully functional protein.

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Polyribosomes and DNA-dependent amino acid incorporation in Escherichia coli extracts

Cited by 17 publications

References 12 publications

An Electron Microscopic Study of Large Bacterial Polyribosomes

An Electron Microscopic Study of Large Bacterial Polyribosomes

Life in science

Monitoring the Progression of Cell-Free Expression of Microbial Rhodopsins by Surface Enhanced IR Spectroscopy: Resolving a Branch Point for Successful/Unsuccessful Folding

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