Structural and functional study of <i>Legionella pneumophila</i> effector RavA

Chung, Ivy Yeuk Wah; Li, Lei; Tyurin, O. V.; Gagarinova, Alla; Wibawa, Raissa; Li, Pengfei; Hartland, Elizabeth L.; Cygler, Mirosław

doi:10.1002/pro.4057

Cited by 8 publications

(2 citation statements)

References 48 publications

(72 reference statements)

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“…It is imperative to confirm that the bait does not autonomously activate reporter genes and is not toxic in Y2HGold in the absence of a prey protein. The pGBKT7 and pGBKT7-AP2N3 plasmids were used in a series of transformation tests to exclude false activation and the toxicity of reporter gene of the system (Chung et al, 2021). The yeast strains Y2HGold and Y187 were transformed using a slightly modified method.…”

Section: Generation Of Yeast Bait Strainmentioning

confidence: 99%

Identification of Interacting Proteins of Transcription Factor DpAP2 Related to Carotenoid Biosynthesis From Marine Microalga Dunaliella parva

Shang

Pang²,

Zhang³

et al. 2022

Front. Mar. Sci.

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Carotenoids are widely distributed and structurally diverse, which have significant roles in the photosynthesis of plants. As a precursor of vitamin A, carotenoids are also antioxidants that reduce various chronic diseases, which are beneficial for human health. Currently, the existing studies concerned the biological roles of APETALA2 (AP2)/ethylene-responsive factor (ERF) genes originated from higher plants. The AP2 superfamily of the transcriptional regulator was identified in higher plants, which was related to growth, development, carotenoid metabolism, and responses to various stresses. However, the regulatory mechanisms of the AP2-modulating carotenoid metabolism have not been reported in microalgae, which remain to be elucidated. Dunaliella parva AP2 (i.e., DpAP2), an important transcription factor, promotes carotenoid accumulation by binding to the promoter of target gene. Here, we identified an important AP2/ERF transcription factor, DpAP2, which could promote carotenoid accumulation by binding to the promoter of target gene. To demonstrate the function of DpAP2, the interacting proteins were identified by the yeast two-hybrid system. The results showed that DpAP2 could interact with three proteins with different activities (DNA-binding transcription factor activity, protein kinase activity, and alpha-D-phosphohexomutase activity); these proteins may be associated with multiple biological processes. This paper laid a good foundation for a deep understanding of the regulatory mechanisms of DpAP2 and genetic engineering breeding in D. parva.

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Section: Generation Of Yeast Bait Strainmentioning

confidence: 99%

Identification of Interacting Proteins of Transcription Factor DpAP2 Related to Carotenoid Biosynthesis From Marine Microalga Dunaliella parva

Shang

Pang²,

Zhang³

et al. 2022

Front. Mar. Sci.

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“…Rab7 is a member of the Ras superfamily and plays a key role in the late stage of autophagy. Therefore, the dysfunction of Rab protein is linked to multiple human diseases, including cancer ( [30][31][32][33][34], infection [30,35,36], cardiac disease [37], and neurodegeneration [38][39][40][41][42][43]. In recent years, an increasing number of researchers [44][45][46][47][48][49][50] have focused on the role of Rab protein in the development of ALS.…”

Section: Introductionmentioning

confidence: 99%

Rab Geranylgeranyltransferase Subunit Beta as a Potential Indicator to Assess the Progression of Amyotrophic Lateral Sclerosis

Yang,

Xin,

Huo

et al. 2023

Brain Sciences

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Background: Currently, there is no effective treatment for amyotrophic lateral sclerosis (ALS), a devastating neurodegenerative disorder. Many biomarkers have been proposed, but because ALS is a clinically heterogeneous disease with an unclear etiology, biomarker discovery for ALS has been challenging due to the lack of specificity of these biomarkers. In recent years, the role of autophagy in the development and treatment of ALS has become a research hotspot. In our previous studies, we found that the expression of RabGGTase (low RABGGTB expression and no change in RABGGTA) is lower in the lumbar and thoracic regions of spinal cord motoneurons in SOD1G93A mice compared with WT (wild-type) mice groups, and upregulation of RABGGTB promoted prenylation modification of Rab7, which promoted autophagy to protect neurons by degrading SOD1. Given that RabGGTase is associated with autophagy and autophagy is associated with inflammation, and based on the above findings, since peripheral blood mononuclear cells are readily available from patients with ALS, we proposed to investigate the expression of RabGGTase in peripheral inflammatory cells. Methods: Information and venous blood were collected from 86 patients diagnosed with ALS between January 2021 and August 2023. Flow cytometry was used to detect the expression of RABGGTB in monocytes from peripheral blood samples collected from patients with ALS and healthy controls. Extracted peripheral blood mononuclear cells (PBMCs) were differentiated in vitro into macrophages, and then the expression of RABGGTB was detected by immunofluorescence. RABGGTB levels in patients with ALS were analyzed to determine their impact on disease progression. Results: Using flow cytometry in monocytes and immunofluorescence in macrophages, we found that RABGGTB expression in the ALS group was significantly higher than in the control group. Age, sex, original location, disease course, C-reactive protein (CRP), and interleukin-6 (IL-6) did not correlate with the ALS functional rating scale—revised (ALSFRS-R), whereas the RABGGTB level was significantly correlated with the ALSFRS-R. In addition, multivariate analysis revealed a significant correlation between RABGGTB and ALSFRS-R score. Further analysis revealed a significant correlation between RABGGTB expression levels and disease progression levels (ΔFS). Conclusions: The RABGGTB level was significantly increased in patients with ALS compared with healthy controls. An elevated RABGGTB level in patients with ALS is associated with the rate of progression in ALS, suggesting that elevated RABGGTB levels in patients with ALS may serve as an indicator for tracking ALS progression.

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Structural and functional study of Legionella pneumophila effector RavA

Chung

Tyurin

et al. 2021

Protein Science

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Legionella pneumophila is an intracellular pathogen that causes Legionnaire's disease in humans. This bacterium can be found in freshwater environments as a free‐living organism, but it is also an intracellular parasite of protozoa. Human infection occurs when inhaled aerosolized pathogen comes into contact with the alveolar mucosa and replicates in alveolar macrophages. Legionella enters the host cell by phagocytosis and redirects the Legionella‐containing phagosomes from the phagocytic maturation pathway. These nascent phagosomes fuse with ER‐derived secretory vesicles and membranes forming the Legionella‐containing vacuole. Legionella subverts many host cellular processes by secreting over 300 effector proteins into the host cell via the Dot/Icm type IV secretion system. The cellular function for many Dot/Icm effectors is still unknown. Here, we present a structural and functional study of L. pneumophila effector RavA (Lpg0008). Structural analysis revealed that the RavA consists of four ~85 residue long α‐helical domains with similar folds, which show only a low level of structural similarity to other protein domains. The ~90 residues long C‐terminal segment is predicted to be natively unfolded. We show that during L. pneumophila infection of human cells, RavA localizes to the Golgi apparatus and to the plasma membrane. The same localization is observed when RavA is expressed in human cells. The localization signal resides within the C‐terminal sequence C409WTSFCGLF417. Yeast‐two‐hybrid screen using RavA as bait identified RAB11A as a potential binding partner. RavA is present in L. pneumophila strains but only distant homologs are found in other Legionella species, where the number of repeats varies.

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Structural and functional study of Legionella pneumophila effector RavA

Cited by 8 publications

References 48 publications

Identification of Interacting Proteins of Transcription Factor DpAP2 Related to Carotenoid Biosynthesis From Marine Microalga Dunaliella parva

Identification of Interacting Proteins of Transcription Factor DpAP2 Related to Carotenoid Biosynthesis From Marine Microalga Dunaliella parva

Rab Geranylgeranyltransferase Subunit Beta as a Potential Indicator to Assess the Progression of Amyotrophic Lateral Sclerosis

Structural and functional study of Legionella pneumophila effector RavA

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