A Short Peptide Designed from Late Embryogenesis Abundant Protein Enhances Acid Tolerance in Escherichia coli

Metwally, Khaled; Ikeno, Shinya

doi:10.1007/s12010-020-03262-5

Cited by 7 publications

(9 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

Section: Discussionmentioning

confidence: 92%

“…The fact that multiple LEA-1 isoforms could improve desiccation tolerance of bacteria turned our attention to conserved motifs contained within these isoforms. Similar motifs from LEA proteins of other organisms have been demonstrated to function in stress resistance [21, 39, 47, 48, 59]. These LEA motifs acquire secondary structure during desiccation-like conditions and specifically are thought to form amphipathic alpha-helices [19, 49, 60].…”

Section: Discussionmentioning

confidence: 99%

“…LEA proteins are commonly found in animals with interesting extremotolerant abilities but with limited genetic tools. Therefore, a common approach has been to study LEA proteins (and fragments thereof) in heterologous systems [20,21,23,45,46,[49][50][51][52]. While such approaches have revealed many potential functions of LEA proteins, there has been limited evidence to bridge protective abilities observed in vitro and in heterologous systems to the endogenous, in vivo context in an animal [10,29,53].…”

Section: Discussionmentioning

confidence: 99%

“…Because all five isoforms tested were sufficient to improve bacterial desiccation resistance, we hypothesized that motifs shared by these isoforms might be sufficient for conferring bacterial desiccation resistance. Group 3 LEA proteins commonly have repeated motifs of 11 amino acids that can protect against stresses including low pH, high salt, and desiccation-induced protein aggregation [8,21,37,[43][44][45][46]. These motifs are predicted to form alpha helices, and circular dichroism of LEA proteins and motif-containing peptides confirms alpha helix formation during conditions when water is limiting [17,18,47].…”

Section: Lea-1 Minimal Motifs Confer Desiccation Resistance To Bacteriamentioning

confidence: 99%

See 3 more Smart Citations

LEA motifs promote desiccation tolerancein vivo

Jd¹,

Goldstein

2021

Preprint

View full text Add to dashboard Cite

Cells and organisms typically cannot survive in the absence of water. However, there are some notable exceptions, including animals such as nematodes, tardigrades, rotifers, and some arthropods. One class of proteins known to play a role in desiccation resistance is the late embryogenesis abundant (LEA) proteins. These largely disordered proteins protect plants and animals alike from desiccation. A multitude of studies have characterized stress-protective functions of LEA proteins in vitro and in heterologous systems. However, the extent to which LEA proteins exhibit the same functions in their native contexts in animals is unclear. Furthermore, nothing is known about the distribution of LEA proteins in multicellular organisms or tissue-specific requirements in conferring stress protection. To study the endogenous function of LEA proteins in an animal, we created a true null mutant of C. elegans LEA-1, as well as endogenous fluorescent reporters of the protein. We confirmed that C. elegans lacking LEA-1 are sensitive to desiccation. LEA-1 mutant animals were also sensitive to heat and osmotic stress and were prone to protein aggregation. During desiccation, LEA-1 expression increased and became more widespread throughout the body. LEA-1 was required at high levels in body wall muscle for animals to survive desiccation and osmotic stress. We identified minimal motifs within LEA-1 that are sufficient to increase desiccation survival of E. coli. Our results provide insights into the endogenous functions and expression dynamics of an LEA protein in a multicellular animal. We show that LEA-1 buffers animals from a broad range of stresses. Our identification of functional motifs within the protein suggests the possibility of engineering LEA-1-derived peptides for desiccation protection.

show abstract

Section: Discussionmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Lea-1 Minimal Motifs Confer Desiccation Resistance To Bacteriamentioning

confidence: 99%

See 2 more Smart Citations

LEA motifs promote desiccation tolerancein vivo

Jd¹,

Goldstein

2021

Preprint

View full text Add to dashboard Cite

show abstract

“…Abiotic or environmental stresses, such as high soil salinity, extreme temperature, water deficiency, and unsuitable pH, are major limiting factors for the growth and productivity of all living organisms [ 33 ]. The group 3 LEA (G3LEA) proteins are well-characterized hydrophilic proteins that upregulate in response to environmental stresses, such as desiccation, freezing, and high salinity [ 34 , 35 ], and they are described not only throughout the plant kingdom but also in other organisms, ranging from invertebrates to prokaryotes [ 36 ].…”

Section: Discussionmentioning

confidence: 99%

Modular Assembly of Ordered Hydrophilic Proteins Improve Salinity Tolerance in Escherichia coli

Guo

Zhao

Tang

et al. 2021

IJMS

View full text Add to dashboard Cite

Most late embryogenesis abundant group 3 (G3LEA) proteins are highly hydrophilic and disordered, which can be transformed into ordered α-helices to play an important role in responding to diverse stresses in numerous organisms. Unlike most G3LEA proteins, DosH derived from Dinococcus radiodurans is a naturally ordered G3LEA protein, and previous studies have found that the N-terminal domain (position 1–103) of DosH protein is the key region for its folding into an ordered secondary structure. Synthetic biology provides the possibility for artificial assembling ordered G3LEA proteins or their analogues. In this report, we used the N-terminal domain of DosH protein as module A (named DS) and the hydrophilic domains (DrHD, BnHD, CeHD, and YlHD) of G3LEA protein from different sources as module B, and artificially assembled four non-natural hydrophilic proteins, named DS + DrHD, DS + BnHD, DS + CeHD, and DS + YlHD, respectively. Circular dichroism showed that the four hydrophile proteins were highly ordered proteins, in which the α-helix contents were DS + DrHD (56.1%), DS + BnHD (53.7%), DS + CeHD (49.1%), and DS + YLHD (64.6%), respectively. Phenotypic analysis showed that the survival rate of recombinant Escherichia coli containing ordered hydrophilic protein was more than 10% after 4 h treatment with 1.5 M NaCl, which was much higher than that of the control group. Meanwhile, in vivo enzyme activity results showed that they had higher activities of superoxide dismutase, catalase, lactate dehydrogenase and less malondialdehyde production. Based on these results, the N-terminal domain of DosH protein can be applied in synthetic biology due to the fact that it can change the order of hydrophilic domains, thus increasing stress resistance.

show abstract