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Spatiotemporal proteomics uncovers cathepsin-dependent macrophage cell death during Salmonella infection.
Nature Microbiology ( IF 28.3 ) Pub Date : 2020-06-08 , DOI: 10.1038/s41564-020-0736-7
Joel Selkrig 1 , Nan Li 1, 2, 3 , Annika Hausmann 4 , Matthew S J Mangan 5, 6 , Matylda Zietek 1 , André Mateus 1 , Jacob Bobonis 1, 7 , Anna Sueki 1, 7 , Haruna Imamura 8 , Bachir El Debs 1, 9 , Gianluca Sigismondo 3 , Bogdan I Florea 10 , Herman S Overkleeft 10 , Nataša Kopitar-Jerala 11 , Boris Turk 11, 12 , Pedro Beltrao 8 , Mikhail M Savitski 1 , Eicke Latz 5, 6, 13 , Wolf-Dietrich Hardt 4 , Jeroen Krijgsveld 3, 14 , Athanasios Typas 1
Affiliation  

The interplay between host and pathogen relies heavily on rapid protein synthesis and accurate protein targeting to ensure pathogen destruction. To gain insight into this dynamic interface, we combined Click chemistry with pulsed stable isotope labelling of amino acids in cell culture to quantify the host proteome response during macrophage infection with the intracellular bacterial pathogen Salmonella enterica Typhimurium. We monitored newly synthesized proteins across different host cell compartments and infection stages. Within this rich resource, we detected aberrant trafficking of lysosomal proteases to the extracellular space and the nucleus. We verified that active cathepsins re-traffic to the nucleus and that these are linked to cell death. Pharmacological cathepsin inhibition and nuclear targeting of a cellular cathepsin inhibitor (stefin B) suppressed S. enterica Typhimurium-induced cell death. We demonstrate that cathepsin activity is required for pyroptotic cell death via the non-canonical inflammasome, and that lipopolysaccharide transfection into the host cytoplasm is sufficient to trigger active cathepsin accumulation in the host nucleus and cathepsin-dependent cell death. Finally, cathepsin inhibition reduced gasdermin D expression, thus revealing an unexpected role for cathepsin activity in non-canonical inflammasome regulation. Overall, our study illustrates how resolution of host proteome dynamics during infection can drive the discovery of biological mechanisms at the host–microbe interface.



中文翻译:

时空蛋白质组学发现沙门氏菌感染期间组织蛋白酶依赖性巨噬细胞死亡。

宿主与病原体之间的相互作用在很大程度上取决于快速的蛋白质合成和准确的蛋白质靶向,以确保病原体的破坏。为了深入了解此动态界面,我们将Click化学与细胞培养物中氨基酸的脉冲稳定同位素标记相结合,以量化在巨噬细胞感染细胞内细菌病原体沙门氏菌的过程中宿主蛋白质组的反应鼠伤寒 我们监测了跨不同宿主细胞区室和感染阶段的新合成蛋白。在这种丰富的资源中,我们检测到溶酶体蛋白酶异常运输到细胞外空间和细胞核。我们验证了活跃的组织蛋白酶重新向细胞核转移,并且这些与细胞死亡有关。细胞组织蛋白酶抑制剂(Stefin B)的药理组织蛋白酶抑制和核靶向作用抑制了肠炎链球菌鼠伤寒引起的细胞死亡。我们证明,组织蛋白酶活性是通过非规范性炎症小体使光热细胞死亡所必需的,脂多糖转染到宿主细胞质中足以​​触发活性组织蛋白酶在宿主细胞核中的积累和组织蛋白酶依赖性细胞死亡。最后,组织蛋白酶抑制降低了胃泌素D的表达,从而揭示了组织蛋白酶活性在非典型的炎症小体调节中的出乎意料的作用。总体而言,我们的研究说明了感染过程中宿主蛋白质组动力学的解决如何推动发现宿主-微生物界面的生物学机制。

更新日期:2020-06-08
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