Dual specificity phosphatase 9: A novel binding partner cum substrate of proapoptotic serine protease HtrA2

Acharya, Saujanya; Dutta, Shubhankar; Mudrale, Snehal Pandav; Bose, Kakoli

doi:10.1016/j.bbrc.2020.09.062

Cited by 6 publications

(11 citation statements)

References 28 publications

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“…Consistent with these observations, our data indicated that HtrA2 regulated microtubule stability by affecting Sirt2 and Hdac6. Intriguingly, α-tubulin was identi ed as the HtrA2 substrate in a proteome-wide screen of Jurkat cell lysates [21]. Moreover, several spindle-related proteins regulate oocyte mitosis or the meiotic process by affecting microtubule acetylation [54].…”

Section: Discussionmentioning

confidence: 99%

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Insufficient HtrA2 causes meiotic defects by inducing cytoskeletal disorganization in aging germinal vesicle oocytes

Gao

Qiu

Cao

et al. 2022

Preprint

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Background High-temperature requirement protease A2 (HtrA2/Omi) is a mitochondrial chaperone highly conserved from bacteria to humans. It plays an important role in mitochondrial homeostasis and apoptosis. In this study, we investigated the role of HtrA2 in mouse oocyte maturation. Methods The role of HtrA2 in mouse oocyte maturation was investigated by employing knockdown (KD) or overexpression (OE) of HtrA2 in young or old GV oocytes. We employed immunoblotting, immunostaining, fluorescent intensity quantification to test the HtrA2 knockdown on the GV oocyte maturation progression, spindle assembly checkpoint, mitochondrial distribution, spindle assembly, chromosome alignment, actin polymerization, DNA damage and chromosome numbers, the level of acetylated tubulin. Results We observed a significant reduction in HtrA2 protein levels in aging germinal vesicle (GV) oocytes. Young oocytes with low levels of HtrA2 due to siRNA knockdown were unable to complete meiosis and were partially blocked at metaphase I (MI). They also displayed significantly more BubR1 on kinetochores, indicating that the spindle assembly checkpoint was triggered at MI. Extrusion of the first polar body (Pb1) was significantly less frequent and oocytes with large polar bodies were observed when HtrA2 was depleted. In addition, HtrA2 knockdown induced meiotic spindle/chromosome disorganization, leading to aneuploidy at metaphase II (MII), possibly due to the elevated level of acetylated tubulin. Importantly, overexpression of HtrA2 partially rescued spindle/chromosome disorganization and reduced the rate of aneuploidy in aging oocytes. Conclusions Collectively, our data suggest that HtrA2 is a key regulator of oocyte maturation, and its deficiency with age appears to contribute to reproduction failure in females.

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Section: Discussionmentioning

confidence: 99%

“…Several cytoskeletal proteins, including α-or β-tubulin, are believed to be HtrA2 substrates based on a proteome-wide screen of Jurkat cell lysates [21]. Spindle assembly depends on the rapid reorganization of dynamic microtubules composed of α-or β-tubulin dimers [22].…”

Section: Introductionmentioning

confidence: 99%

Insufficient HtrA2 causes meiotic defects by inducing cytoskeletal disorganization in aging germinal vesicle oocytes

Gao

Qiu

Cao

et al. 2022

Preprint

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show abstract

“…Consistent with these observations, our data indicated that HtrA2 may regulate microtubule stability by affecting both Sirt2 and Hdac6 expression. Intriguingly, α-tubulin was identified as the HtrA2 substrate in a proteome-wide screen of Jurkat cell lysates [ 21 ]. Moreover, several spindle-related proteins regulate oocyte mitosis or the meiotic process by affecting microtubule acetylation [ 56 ].…”

Section: Discussionmentioning

confidence: 99%

“…Several cytoskeletal proteins, including α- or β-tubulin, are believed to be HtrA2 substrates based on a proteome-wide screen of Jurkat cell lysates [ 21 ]. Spindle organization depends on the rapid reorganization of dynamic microtubules composed of α- or β-tubulin dimers [ 22 ].…”

Section: Introductionmentioning

confidence: 99%

Insufficient HtrA2 causes meiotic defects in aging germinal vesicle oocytes

Gao

Qiu

Cao

et al. 2022

Reprod Biol Endocrinol

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Background High-temperature requirement protease A2 (HtrA2/Omi) is a mitochondrial chaperone that is highly conserved from bacteria to humans. It plays an important role in mitochondrial homeostasis and apoptosis. In this study, we investigated the role of HtrA2 in mouse oocyte maturation. Methods The role of HtrA2 in mouse oocyte maturation was investigated by employing knockdown (KD) or overexpression (OE) of HtrA2 in young or old germinal vesicle (GV) oocytes. We employed immunoblotting, immunostaining, fluorescent intensity quantification to test the HtrA2 knockdown on the GV oocyte maturation progression, spindle assembly checkpoint, mitochondrial distribution, spindle organization, chromosome alignment, actin polymerization, DNA damage and chromosome numbers and acetylated tubulin levels. Results We observed a significant reduction in HtrA2 protein levels in aging germinal vesicle (GV) oocytes. Young oocytes with low levels of HtrA2 due to siRNA knockdown were unable to complete meiosis and were partially blocked at metaphase I (MI). They also displayed significantly more BubR1 on kinetochores, indicating that the spindle assembly checkpoint was triggered at MI. Extrusion of the first polar body (Pb1) was significantly less frequent and oocytes with large polar bodies were observed when HtrA2 was depleted. In addition, HtrA2 knockdown induced meiotic spindle/chromosome disorganization, leading to aneuploidy at metaphase II (MII), possibly due to the elevated level of acetylated tubulin. Importantly, overexpression of HtrA2 partially rescued spindle/chromosome disorganization and reduced the rate of aneuploidy in aging GV oocytes. Conclusions Collectively, our data suggest that HtrA2 is a key regulator of oocyte maturation, and its deficiency with age appears to contribute to reproduction failure in females.

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“…This multitiered regulation of HtrA2 activation might be critical toward prevention of untimely proteolysis as well as accurately controlling its involvement in different pathophysiological pathways such as apoptosis, protein quality-control, cancer, arthritis, and neurodegeneration, where it cleaves a wide spectrum of substrates in different subcellular locations. This is substantiated by the identification and characterization of protein-protein interactions involving HtrA2 and its substrates such as Inhibitor of Apoptosis Proteins (IAPs), hematopoietic cell-specific protein-1-(HS1)-associated protein X-1 (Hax-1), Dual-specificity phosphatase-9 (DUSP-9), a gene associated with retinoic and interferon-induced mortality-19 protein (GRIM-19) and Phosphoprotein enriched in astrocytes-15 (Pea-15) (Chaganti et al, 2019;Acharya et al, 2020;Kummari et al, 2021) that unlike other HtrAs are interestingly not restricted to the C-terminal PDZ domains. The holistic enumeration of HtrA2's activation network has been vividly illustrated in Figure 2 and the mechanism is elaborated in the figure legend.…”

Section: Active Site Conformation and Multiple Activation Mechanisms ...mentioning

confidence: 99%

Unraveling the Dichotomy of Enigmatic Serine Protease HtrA2

Chakraborty

Bose

2022

Front. Mol. Biosci.

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Mitochondrial high-temperature requirement protease A2 (HtrA2) is an integral member of the HtrA family of serine proteases that are evolutionarily conserved from prokaryotes to humans. Involvement in manifold intricate cellular networks and diverse pathophysiological functions make HtrA2 the most enigmatic moonlighting protease amongst the human HtrAs. Despite perpetuating the oligomeric architecture and overall structural fold of its homologs that comprises serine protease and regulatory PDZ domains, subtle conformational alterations and dynamic enzymatic regulation through the distinct allosteric mode of action lead to its functional diversity. This mitochondrial protease upon maturation, exposes its one-of-a-kind N-terminal tetrapeptide (AVPS) motif that binds and subsequently cleaves Inhibitor of Apoptosis Proteins (IAPs) thus promoting cell death, and posing as an important molecule for therapeutic intervention. Interestingly, unlike its other human counterparts, HtrA2 has also been implicated in maintaining the mitochondrial integrity through a bi-functional chaperone-protease activity, the on-off switch of which is yet to be identified. Furthermore, its ability to activate a wide repertoire of substrates through both its N- and C-terminal regions presumably has calibrated its association with several cellular pathways and hence diseases including neurodegenerative disorders and cancer. Therefore, the exclusive structural attributes of HtrA2 that involve multimodal activation, intermolecular PDZ-protease crosstalk, and an allosterically-modulated trimeric active-site ensemble have enabled the protease to evolve across species and partake functions that are fine-tuned for maintaining cellular homeostasis and mitochondrial proteome quality control in humans. These unique features along with its multitasking potential make HtrA2 a promising therapeutic target both in cancer and neurodegeneration.

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Dual specificity phosphatase 9: A novel binding partner cum substrate of proapoptotic serine protease HtrA2

Cited by 6 publications

References 28 publications

Insufficient HtrA2 causes meiotic defects by inducing cytoskeletal disorganization in aging germinal vesicle oocytes

Insufficient HtrA2 causes meiotic defects by inducing cytoskeletal disorganization in aging germinal vesicle oocytes

Insufficient HtrA2 causes meiotic defects in aging germinal vesicle oocytes

Unraveling the Dichotomy of Enigmatic Serine Protease HtrA2

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