A Trapping Approach Reveals Novel Substrates and Physiological Functions of the Essential Protease FtsH in Escherichia coli

Westphal, Kai; Langklotz, Sina; Thomanek, Nikolas; Narberhaus, Franz

doi:10.1074/jbc.m112.388470

Cited by 68 publications

(101 citation statements)

References 69 publications

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“…Thus far, conditional proteolysis has only been reported for a few substrates. For instance, (i) increasing temperatures trigger the degradation of the homoserine transsuccinylase HTS (MetA), which catalyzes the first step in the de novo methionine biosynthesis and thereby affects the rate of translation (18), (ii) the stability of the DNA-binding replication inhibitor CspD is adjusted according to the growth rate and growth phase of E. coli cells (19), (iii) anti-sigma factor guided degradation controls the amount of the stationary phase sigma factor RpoS (33), and (iv) the formate dehydrogenase subunit FdoH and the uncharacterized YfgM protein are degraded by FtsH in response to various growth conditions (12).…”

Section: Discussionmentioning

confidence: 99%

“…The C terminus of LpxC has been shown to contain a nonpolar, sequence-and length-specific signal for degradation by the FtsH protease (6,7). Besides its essential role in controlling LPS biosynthesis, FtsH is involved in the quality control of membrane proteins and the regulation of various cellular mechanisms, in particular under stress conditions (12)(13)(14)(15). Although about 15 different FtsH substrates are known, there are many open questions concerning their precise degradation mechanism.…”

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confidence: 99%

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FtsH-Mediated Coordination of Lipopolysaccharide Biosynthesis in Escherichia coli Correlates with the Growth Rate and the Alarmone (p)ppGpp

2013

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The outer membrane is the first line of defense for Gram-negative bacteria and serves as a major barrier for antibiotics and other harmful substances. The biosynthesis of lipopolysaccharides (LPS), the essential component of the outer membrane, must be tightly controlled as both too much and too little LPS are toxic. In Escherichia coli, the cellular level of the key enzyme LpxC, which catalyzes the first committed step in LPS biosynthesis, is adjusted by proteolysis carried out by the essential and membrane-bound protease FtsH. Here, we demonstrate that LpxC is degraded in a growth rate-dependent manner with half-lives between 4 min and >2 h. According to the cellular demand for LPS biosynthesis, LpxC is degraded during slow growth but stabilized when cells grow rapidly. Disturbing the balance between LPS and phospholipid biosynthesis in favor of phospholipid production in an E. coli strain encoding a hyperactive FabZ protein abolishes growth rate dependency of LpxC proteolysis. Lack of the alternative sigma factor RpoS or inorganic polyphosphates, which are known to mediate growth rate-dependent gene regulation in E. coli, did not affect proteolysis of LpxC. In contrast, absence of RelA and SpoT, which synthesize the alarmone (p)ppGpp, deregulated LpxC degradation resulting in rapid proteolysis in fast-growing cells and stabilization during slow growth. Our data provide new insights into the essential control of LPS biosynthesis in E. coli.

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Section: Discussionmentioning

confidence: 99%

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FtsH-Mediated Coordination of Lipopolysaccharide Biosynthesis in Escherichia coli Correlates with the Growth Rate and the Alarmone (p)ppGpp

2013

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“…Comprehensive substrate identification is necessary for determination of a common sequence signature or regulatory motif. Using substrate-trapping approaches, multiple substrates for Clp and FtsH proteases have been isolated in bacteria and in mammalian mitochondria Neher et al, 2003;Westphal et al, 2012;Bhat et al, 2013;Feng et al, 2013;Graham et al, 2013;Bittner et al, 2015). Trapping strategies use mutated, inactive forms of the protease or chaperone domains.…”

Section: Proteolytic Remodeling Of the Photosynthetic Machineries In mentioning

confidence: 99%

Chloroplast Proteases: Updates on Proteolysis within and across Suborganellar Compartments

2016

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ORCID IDs: 0000-0003-1718-9121 (Y.K.); 0000-0001-9747-5042 (W.S.).Chloroplasts originated from the endosymbiosis of ancestral cyanobacteria and maintain transcription and translation machineries for around 100 proteins. Most endosymbiont genes, however, have been transferred to the host nucleus, and the majority of the chloroplast proteome is composed of nucleus-encoded proteins that are biosynthesized in the cytosol and then imported into chloroplasts. How chloroplasts and the nucleus communicate to control the plastid proteome remains an important question. Protein-degrading machineries play key roles in chloroplast proteome biogenesis, remodeling, and maintenance. Research in the past few decades has revealed more than 20 chloroplast proteases, which are localized to specific suborganellar locations. In particular, two energy-dependent processive proteases of bacterial origin, Clp and FtsH, are central to protein homeostasis. Processing endopeptidases such as stromal processing peptidase and thylakoidal processing peptidase are involved in the maturation of precursor proteins imported into chloroplasts by cleaving off the amino-terminal transit peptides. Presequence peptidases and organellar oligopeptidase subsequently degrade the cleaved targeting peptides. Recent findings have indicated that not only intraplastidic but also extraplastidic processive protein-degrading systems participate in the regulation and quality control of protein translocation across the envelopes. In this review, we summarize current knowledge of the major chloroplast proteases in terms of type, suborganellar localization, and diversification. We present details of these degradation processes as case studies according to suborganellar compartment (envelope, stroma, and thylakoids). Key questions and future directions in this field are discussed.Over 1 billion years of plastid evolution since the endosymbiosis of ancestral cyanobacteria (Douzery et al., 2004), chloroplast biogenesis has gained complexity, with large sets of the endosymbiont genes being transferred to host nuclear genomes. While only 100 endosymbiont genes remain in the plastid genome, with the corresponding proteins biosynthesized there, the nuclear genes have often gained complexity by duplication and diversification of the original endosymbiotic genes. This complexity raises numerous questions regarding (1) at what level gene expression is coordinately controlled, (2) what molecules coordinate the cross talk between chloroplasts and the nucleus, (3) how proteins get across membranes and become imported into chloroplasts, and (4) how the stoichiometries of nucleus-and chloroplast-encoded subunits within individual chloroplast protein complexes such as photosystems and Rubisco are strictly maintained.Given these questions, chloroplast biogenesis has remained a central subject in plant physiology for the last few decades (Jarvis and López-Juez, 2013). In our view, the aforementioned questions point to the importance of protein homeostasis and posttranslational modificat...

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“…In order to abolish the proteolytic activity of FTSH4 protease, a highly conserved histidine at the position 486 (a Zn 2+ -binding site, in the HExxH motif ) was mutated into a tyrosine residue (Westphal et al, 2012). The mutation was introduced on FTSH4.FLAG pENTR plasmid using a QuikChange Lightening site-directed mutagenesis kit (Agilent Technologies) and primers Mut_prot_H_FP and Mut_prot_H_RP (Table S2) resulting in a FTSH4(HIS).FLAG pENTR plasmid.…”

Section: Generation Of the Ftsh4-1 Ftsh4(h486y) Mutant Linementioning

confidence: 99%

The plant i-AAA protease controls the turnover of an essential mitochondrial protein import component

Opalińska

Parys

Murcha

et al. 2018

Journal of Cell Science

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Mitochondria are multifunctional organelles that play a central role in energy metabolism. Owing to the life-essential functions of these organelles, mitochondrial content, quality and dynamics are tightly controlled. Across the species, highly conserved ATP-dependent proteases prevent malfunction of mitochondria through versatile activities. This study focuses on a molecular function of the plant mitochondrial inner membrane-embedded AAA protease (denoted i-AAA) FTSH4, providing its first bona fide substrate. Here, we report that the abundance of the Tim17-2 protein, an essential component of the TIM17:23 translocase (Tim17-2 together with Tim50 and Tim23), is directly controlled by the proteolytic activity of FTSH4. Plants that are lacking functional FTSH4 protease are characterized by significantly enhanced capacity of preprotein import through the TIM17:23-dependent pathway. Taken together, with the observation that FTSH4 prevents accumulation of Tim17-2, our data point towards the role of this i-AAA protease in the regulation of mitochondrial biogenesis in plants.

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A Trapping Approach Reveals Novel Substrates and Physiological Functions of the Essential Protease FtsH in Escherichia coli

Cited by 68 publications

References 69 publications

FtsH-Mediated Coordination of Lipopolysaccharide Biosynthesis in Escherichia coli Correlates with the Growth Rate and the Alarmone (p)ppGpp

FtsH-Mediated Coordination of Lipopolysaccharide Biosynthesis in Escherichia coli Correlates with the Growth Rate and the Alarmone (p)ppGpp

Chloroplast Proteases: Updates on Proteolysis within and across Suborganellar Compartments

The plant i-AAA protease controls the turnover of an essential mitochondrial protein import component

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