A myeloid leukemia factor homolog is involved in tolerance to stresses and stress-induced protein metabolism in Giardia lamblia
Abstract Background The eukaryotic membrane vesicles contain specific sets of proteins that determine vesicle function and shuttle with specific destination. Giardia lamblia contains unknown cytosolic vesicles that are related to the identification of a homolog of human myeloid leukemia factor (MLF)...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-04-01
|
| Series: | Biology Direct |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13062-023-00378-6 |
| _version_ | 1797836581797625856 |
|---|---|
| author | Jui-Hsuan Wu Jen-Chi Lee Chun-Che Ho Pei-Wei Chiu Chin-Hung Sun |
| author_facet | Jui-Hsuan Wu Jen-Chi Lee Chun-Che Ho Pei-Wei Chiu Chin-Hung Sun |
| author_sort | Jui-Hsuan Wu |
| collection | DOAJ |
| description | Abstract Background The eukaryotic membrane vesicles contain specific sets of proteins that determine vesicle function and shuttle with specific destination. Giardia lamblia contains unknown cytosolic vesicles that are related to the identification of a homolog of human myeloid leukemia factor (MLF) named MLF vesicles (MLFVs). Previous studies suggest that MLF also colocalized with two autophagy machineries, FYVE and ATG8-like protein, and that MLFVs are stress-induced compartments for substrates of the proteasome or autophagy in response to rapamycin, MG132, and chloroquine treatment. A mutant protein of cyclin-dependent kinase 2, CDK2m3, was used to understand whether the aberrant proteins are targeted to degradative compratments. Interestingly, MLF was upregulated by CDK2m3 and they both colocalized within the same vesicles. Autophagy is a self-digestion process that is activated to remove damaged proteins for preventing cell death in response to various stresses. Because of the absence of some autophagy machineries, the mechanism of autophagy is unclear in G. lamblia. Results In this study, we tested the six autophagosome and stress inducers in mammalian cells, including MG132, rapamycin, chloroquine, nocodazole, DTT, and G418, and found that their treatment increased reactive oxygen species production and vesicle number and level of MLF, FYVE, and ATG8-like protein in G. lamblia. Five stress inducers also increased the CDK2m3 protein levels and vesicles. Using stress inducers and knockdown system for MLF, we identified that stress induction of CDK2m3 was positively regulated by MLF. An autophagosome-reducing agent, 3-methyl adenine, can reduce MLF and CDK2m3 vesicles and proteins. In addition, knockdown of MLF with CRISPR/Cas9 system reduced cell survival upon treatment with stress inducers. Our newly developed complementation system for CRISPR/Cas9 indicated that complementation of MLF restored cell survival in response to stress inducers. Furthermore, human MLF2, like Giardia MLF, can increase cyst wall protein expression and cyst formation in G. lamblia, and it can colocalize with MLFVs and interact with MLF. Conclusions Our results suggest that MLF family proteins are functionally conserved in evolution. Our results also suggest an important role of MLF in survival in stress conditions and that MLFVs share similar stress-induced characteristics with autophagy compartments. |
| first_indexed | 2024-04-09T15:12:21Z |
| format | Article |
| id | doaj.art-e59e2cbecf7f4dc887c5b31e01f10e9b |
| institution | Directory Open Access Journal |
| issn | 1745-6150 |
| language | English |
| last_indexed | 2024-04-09T15:12:21Z |
| publishDate | 2023-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Biology Direct |
| spelling | doaj.art-e59e2cbecf7f4dc887c5b31e01f10e9b2023-04-30T11:09:57ZengBMCBiology Direct1745-61502023-04-0118112010.1186/s13062-023-00378-6A myeloid leukemia factor homolog is involved in tolerance to stresses and stress-induced protein metabolism in Giardia lambliaJui-Hsuan Wu0Jen-Chi Lee1Chun-Che Ho2Pei-Wei Chiu3Chin-Hung Sun4Department of Tropical Medicine and Parasitology, College of Medicine, National Taiwan UniversityDepartment of Tropical Medicine and Parasitology, College of Medicine, National Taiwan UniversityDepartment of Tropical Medicine and Parasitology, College of Medicine, National Taiwan UniversityDepartment of Tropical Medicine and Parasitology, College of Medicine, National Taiwan UniversityDepartment of Tropical Medicine and Parasitology, College of Medicine, National Taiwan UniversityAbstract Background The eukaryotic membrane vesicles contain specific sets of proteins that determine vesicle function and shuttle with specific destination. Giardia lamblia contains unknown cytosolic vesicles that are related to the identification of a homolog of human myeloid leukemia factor (MLF) named MLF vesicles (MLFVs). Previous studies suggest that MLF also colocalized with two autophagy machineries, FYVE and ATG8-like protein, and that MLFVs are stress-induced compartments for substrates of the proteasome or autophagy in response to rapamycin, MG132, and chloroquine treatment. A mutant protein of cyclin-dependent kinase 2, CDK2m3, was used to understand whether the aberrant proteins are targeted to degradative compratments. Interestingly, MLF was upregulated by CDK2m3 and they both colocalized within the same vesicles. Autophagy is a self-digestion process that is activated to remove damaged proteins for preventing cell death in response to various stresses. Because of the absence of some autophagy machineries, the mechanism of autophagy is unclear in G. lamblia. Results In this study, we tested the six autophagosome and stress inducers in mammalian cells, including MG132, rapamycin, chloroquine, nocodazole, DTT, and G418, and found that their treatment increased reactive oxygen species production and vesicle number and level of MLF, FYVE, and ATG8-like protein in G. lamblia. Five stress inducers also increased the CDK2m3 protein levels and vesicles. Using stress inducers and knockdown system for MLF, we identified that stress induction of CDK2m3 was positively regulated by MLF. An autophagosome-reducing agent, 3-methyl adenine, can reduce MLF and CDK2m3 vesicles and proteins. In addition, knockdown of MLF with CRISPR/Cas9 system reduced cell survival upon treatment with stress inducers. Our newly developed complementation system for CRISPR/Cas9 indicated that complementation of MLF restored cell survival in response to stress inducers. Furthermore, human MLF2, like Giardia MLF, can increase cyst wall protein expression and cyst formation in G. lamblia, and it can colocalize with MLFVs and interact with MLF. Conclusions Our results suggest that MLF family proteins are functionally conserved in evolution. Our results also suggest an important role of MLF in survival in stress conditions and that MLFVs share similar stress-induced characteristics with autophagy compartments.https://doi.org/10.1186/s13062-023-00378-6CystMLFCDK2 mutantGiardiaDifferentiationNocodazole |
| spellingShingle | Jui-Hsuan Wu Jen-Chi Lee Chun-Che Ho Pei-Wei Chiu Chin-Hung Sun A myeloid leukemia factor homolog is involved in tolerance to stresses and stress-induced protein metabolism in Giardia lamblia Biology Direct Cyst MLF CDK2 mutant Giardia Differentiation Nocodazole |
| title | A myeloid leukemia factor homolog is involved in tolerance to stresses and stress-induced protein metabolism in Giardia lamblia |
| title_full | A myeloid leukemia factor homolog is involved in tolerance to stresses and stress-induced protein metabolism in Giardia lamblia |
| title_fullStr | A myeloid leukemia factor homolog is involved in tolerance to stresses and stress-induced protein metabolism in Giardia lamblia |
| title_full_unstemmed | A myeloid leukemia factor homolog is involved in tolerance to stresses and stress-induced protein metabolism in Giardia lamblia |
| title_short | A myeloid leukemia factor homolog is involved in tolerance to stresses and stress-induced protein metabolism in Giardia lamblia |
| title_sort | myeloid leukemia factor homolog is involved in tolerance to stresses and stress induced protein metabolism in giardia lamblia |
| topic | Cyst MLF CDK2 mutant Giardia Differentiation Nocodazole |
| url | https://doi.org/10.1186/s13062-023-00378-6 |
| work_keys_str_mv | AT juihsuanwu amyeloidleukemiafactorhomologisinvolvedintolerancetostressesandstressinducedproteinmetabolismingiardialamblia AT jenchilee amyeloidleukemiafactorhomologisinvolvedintolerancetostressesandstressinducedproteinmetabolismingiardialamblia AT chuncheho amyeloidleukemiafactorhomologisinvolvedintolerancetostressesandstressinducedproteinmetabolismingiardialamblia AT peiweichiu amyeloidleukemiafactorhomologisinvolvedintolerancetostressesandstressinducedproteinmetabolismingiardialamblia AT chinhungsun amyeloidleukemiafactorhomologisinvolvedintolerancetostressesandstressinducedproteinmetabolismingiardialamblia AT juihsuanwu myeloidleukemiafactorhomologisinvolvedintolerancetostressesandstressinducedproteinmetabolismingiardialamblia AT jenchilee myeloidleukemiafactorhomologisinvolvedintolerancetostressesandstressinducedproteinmetabolismingiardialamblia AT chuncheho myeloidleukemiafactorhomologisinvolvedintolerancetostressesandstressinducedproteinmetabolismingiardialamblia AT peiweichiu myeloidleukemiafactorhomologisinvolvedintolerancetostressesandstressinducedproteinmetabolismingiardialamblia AT chinhungsun myeloidleukemiafactorhomologisinvolvedintolerancetostressesandstressinducedproteinmetabolismingiardialamblia |