Tubulin Tail Sequences and Post-translational Modifications Regulate Closure of Mitochondrial Voltage-dependent Anion Channel (VDAC)

Sheldon, Kely L.; Gurnev, Philip A.; Bezrukov, Sergey M.; Sackett, Dan L.

doi:10.1074/jbc.m115.678854

Cited by 29 publications

(27 citation statements)

References 26 publications

(19 reference statements)

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“…The on-rate strongly depends on the -tubulin subunit domain membrane-binding efficiency (38,39). In this respect, the obtained results expectedly differ from those of a recent study of VDAC blockage by CTT-albumin chimeras synthesized using the methods of click chemistry (46). The major difference between these two constructsalbumin-based and yeast tubulin-based -is that water-soluble albumin does not bind to the membrane surfaces and, therefore, does not belong to the class of peripheral membrane proteins (47,48).…”

Section: A Model Of the Tubulin Ctt-vdac Interaction -contrasting

confidence: 73%

Sequence diversity of tubulin isotypes in regulation of the mitochondrial voltage-dependent anion channel

Rostovtseva

Gurnev

Hoogerheide

et al. 2018

Journal of Biological Chemistry

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The microtubule protein tubulin is a heterodimer comprising / subunits, in which each subunit features multiple isotypes in vertebrates. For example, seven -tubulin and eight -tubulin isotypes in the human tubulin gene family vary mostly in the length and primary sequence of the disordered anionic C-terminal tails (CTTs). The biological reason for such sequence diversity remains a topic of vigorous enquiry. Here, we demonstrate that it may be a key feature of tubulin's role in regulation of the permeability of the mitochondrial outer membrane voltagedependent anion channel (VDAC). Using recombinant yeast /-tubulin constructs with -CTTs, -CTTs, or both from various human tubulin isotypes, we probed their interactions with VDAC reconstituted into planar lipid bilayers. A comparative study of the blockage kinetics revealed that either -CTTs or -CTTs block VDAC pore and that the efficiency of blockage by individual CTTs spans two orders of magnitude, depending on the CTT isotype.-Tubulin constructs, notably 3, blocked VDAC most effectively. We quantitatively describe these experimental results using a physical model that accounts only for the number and distribution of charges in the CTT, and not for the interactions between specific residues on the CTT and VDAC pore. Based on these results, we speculate that the effectiveness of VDAC regulation by tubulin depends on the predominant tubulin isotype in a cell. Consequently, the fluxes of ATP/ADP http://www.jbc.org/cgi

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Section: A Model Of the Tubulin Ctt-vdac Interaction -contrasting

confidence: 73%

Sequence diversity of tubulin isotypes in regulation of the mitochondrial voltage-dependent anion channel

Rostovtseva

Gurnev

Hoogerheide

et al. 2018

Journal of Biological Chemistry

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“…An intriguing possible explanation for both the apparent paradoxes is that soluble tubulin dimer has some function other than building microtubules, which necessitates tight control of its concentration. For example, soluble tubulin was proposed to gate Voltage-Dependent Anion Channels (VDAC) in mitochondria 38 , and it is conceivable that autoregulation evolved to regulate this function, not microtubule assembly. Another possibility is that autoregulation evolved to balance the synthesis of α-and β-tubulin subunits, in which case other components of the microtubule cytoskeleton are irrelevant.…”

Section: Autoregulation Suggests Functions Of Tubulin Beyond Buildingmentioning

confidence: 99%

Tubulin mRNA stability is sensitive to change in microtubule dynamics caused by multiple physiological and toxic cues

Gasic

Boswell

Mitchison

2019

Preprint

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The localization, mass, and dynamics of microtubules are important in many processes. Cells may actively monitor the state of their microtubules and respond to perturbation, but how this occurs outside mitosis is poorly understood. We used gene expression analysis in quiescent cells to analyze responses to subtle and strong perturbation of microtubules. Genes encoding α-, β, and -tubulins, but not -or -tubulins, exhibited the strongest differential expression response to microtubule-stabilizing versus destabilizing drugs. Q-PCR of exon versus intron sequences confirmed that these changes were caused by regulation of tubulin mRNA stability and not transcription. Using tubulin mRNA stability as a signature to query the GEO database, we find that tubulin genes respond to toxins known to damage microtubules. Importantly, we find many other experimental perturbations, including multiple signaling and metabolic inputs that trigger tubulin differential expression, suggesting their novel role in the regulation of microtubule cytoskeleton. Mechanistic follow up confirms that one important physiological signal, phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) activity, indeed regulates tubulin mRNA stability via changes in microtubule dynamics. We propose that that tubulin gene expression is regulated as part of many coordinated biological responses, with wide implications in physiology and toxicology. Furthermore, we present a new way to discover microtubule regulation using genomics.

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“…Chimeric protein constructs of BSA and tubulin CTT peptides were produced using chemical linkage via aromatic-hydrazine-aromatic-aldehyde click chemistry (28). Peptides corresponding to the CTT sequences of human b-tubulin (TUBB, NP_821133; -ATAEEEEDFGEEAEEEA), human a-tubulin (TUBA1A, NP_001257328; -DSVEGEGEGEEEGEEY), and detyrosinated a-tubulin (-DSVEGEGEGEEEGEE) were obtained from Peptide 2.0 (Chantilly, VA).…”

Section: Ctt Peptide and Bsa-ctt Chimerasmentioning

confidence: 99%

“…To address these questions, we created peptide sequences of human aand b-tubulin CTTs. Using click chemistry (43), we covalently bound the CTT constructs to BSA protein to make CTT-BSA chimeras (see Materials and Methods and Sheldon et al (28)). A background concentration of BSA protein already exists in our assays, so the addition of BSA is controlled.…”

Section: Biophysical Journal 109(12) 2546-2561mentioning

confidence: 99%

Katanin Severing and Binding Microtubules Are Inhibited by Tubulin Carboxy Tails

Bailey

Sackett

Ross

2015

Biophysical Journal

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Microtubule dynamics in cells are regulated by associated proteins that can be either stabilizers or destabilizers. A class of destabilizers that is important in a large number of cellular activities is the microtubule-severing enzymes, yet little is known about how they function. Katanin p60 was the first ATPase associated with microtubule severing. Here, we investigate the activity of katanin severing using a GFP-labeled human version. We quantify the effect of katanin concentration on katanin binding and severing activity. We find that free tubulin can inhibit severing activity by interfering with katanin binding to microtubules. The inhibition is mediated by the sequence of the tubulin and specifically depends on the carboxy-terminal tails. We directly investigate the inhibition effect of tubulin carboxy-terminal tails using peptide sequences of α-, β-, or detyrosinated α-tubulin tails that have been covalently linked to bovine serum albumin. Our results show that β-tubulin tails are the most effective at inhibiting severing, and that detyrosinated α-tubulin tails are the least effective. These results are distinct from those for other severing enzymes and suggest a scheme for regulation of katanin activity in cells dependent on free tubulin concentration and the modification state of the tubulin.

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Tubulin Tail Sequences and Post-translational Modifications Regulate Closure of Mitochondrial Voltage-dependent Anion Channel (VDAC)

Cited by 29 publications

References 26 publications

Sequence diversity of tubulin isotypes in regulation of the mitochondrial voltage-dependent anion channel

Sequence diversity of tubulin isotypes in regulation of the mitochondrial voltage-dependent anion channel

Tubulin mRNA stability is sensitive to change in microtubule dynamics caused by multiple physiological and toxic cues

Katanin Severing and Binding Microtubules Are Inhibited by Tubulin Carboxy Tails

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