Mini Review : Biochemical and molecular features of LRRK2 and its pathophysiological roles in Parkinson`s disease

( Wongi Seol ) 한국생화학분자생물학회 (구 한국생화학회) 2010 BMB Reports Vol.43 No.4

Parkinson`s disease (PD) is the second most common neurodegenerative disease, and 5-10% of the PD cases are genetically inherited as familial PD (FPD). LRRK2 (leucine-rich repeat kinase 2) was first reported in 2004 as a gene corresponding to PARK8, an autosomal gene whose dominant mutations cause familial PD. LRRK2 contains both active kinase and GTPase domains as well as protein-protein interaction motifs such as LRR (leucine-rich repeat) and WD40. Most pathogenic LRRK2 mutations are located in either the GTPase or kinase domain, implying important roles for the enzymatic activities in PD pathogenic mechanisms. In comparison to other PD causative genes such as parkin and PINK1, LRRK2 exhibits two important features. One is that LRRK2`s mutations (especially the G2019S mutation) were observed in sporadic as well as familial PD patients. Another is that, among the various PD-causing genes, pathological characteristics observed in patients carrying LRRK2 mutations are the most similar to patients with sporadic PD. Because of these two observations, LRRK2 has been intensively investigated for its pathogenic mechanism (s) and as a target gene for PD therapeutics. In this review, the general biochemical and molecular features of LRRK2, the recent results of LRRK2 studies and LRRK2`s therapeutic potential as a PD target gene will be discussed. [BMB reports 2010; 43(4): 233-244]

Identification and Characterization of IL2/15Rβ and Its Isoforms in Ducks

Jip seol Jeong,Woo H Kim,Cherry P. Fernandez,Youn Jeong Lee,Wongi Min 대한수의학회 2013 대한수의학회 학술대회발표집 Vol.2013 No.-

T0901317 as an Inhibitor of Transcriptional Activation of Constitutive Androstane Receptor (CAR)

Hyun-Ha Kim(김현하),Wongi Seol(설원기) 한국생명과학회 2011 생명과학회지 Vol.21 No.4

T0901317은 핵수용체 전사인자인 liver X receptor (LXR, NR1H2/3)의 강력한 합성 리간드이다. 그러나, T0901317은 farnesoid X receptor (FXR, NR1H4)와 pregnane X receptor (PXR, NR1I2)에 대해 작용물질(agonist)로, androgen receptor (AR, NR3C4)와 rertinoid-related orphan receptor-α (ROR-α, NR1F1)에 대해 길항제(antagonist)로 작용하여, LXR외에 적어도 다른 4종의 핵수용체에 대해 그 활성을 조절한다고 보고되었다. 우리는 T0901317이 또 다른 핵수용체인 constitutive androstane receptor (CAR, NR1I3)에 대해 저해제로 기능함을 확인하였다. CAR는 이미 T0901317에 의해 기능이 조절된다고 알려진 PXR, FXR, LXR과 더불어 간에서 생체이물과 콜레스테롤의 대사작용에 중요한 역할을 하므로 T0901317에 의해 CAR의 활성이 조절된다는 사실은, 간세포에서 T0901317을 이용한 실험결과를 해석할 때 세포 내에 이미 존재하는 이들 핵수용체 단백질의 영향을 고려하여 주의깊게 해석해야 함을 의미한다. T0901317 is a potent synthetic ligand for liver X receptor (LXR, NR1H2/3), a member of the nuclear receptor superfamily that functions as a transcription factor. However, T0901317 has been also reported to modulate the activity at least four other nuclear receptors (NRs), acting as agonists for farnesoid X receptor (FXR, NR1H4) and pregnane X receptor (PXR, NR1I2) and as antagonists for androgen receptor (AR, NR3C4) and retinoid-related orphan receptor-α (ROR-α, NR1F1). We report here that T0901317 can also function as an inhibitor for constitutive androstane receptor (CAR, NR1I3). Since CAR is a major player of xenobiotic and cholesterol metabolism in the liver, along with PXR, FXR and LXR, which are reported to be regulated by T0901317, this further complicates the interpretation of potential results with T0901317 in liver cells.

Effect of leucine-rich repeat kinase 2 (LRRK2) on protein synthesis

Kim, Hyejung,Son, Ilhong,Seol, Wongi The Korean Society for Integrative Biology 2018 Animal cells and systems Vol.22 No.1

Mutations in the leucine-rich repeat kinase 2 (LRRK2) cause Parkinson's disease (PD) in an autosomal dominant manner. Pathogenic mutations of LRRK2 such as G2019S and R1441C have been observed as common genetic causes of PD. Recently, LRRK2 has been reported to increase the reporter protein synthesis in both cap-dependent and -independent manners via phosphorylation of the ribosomal protein RPS15. In this study, we tested whether LRRK2 recombinant protein would directly increase protein synthesis using a well-defined in vitro coupled transcription/translation system. Addition of commercial full-length LRRK2 or GST-fused N-terminal-deleted LRRK2 recombinant proteins to the system showed no change of protein synthesis, as measured by luciferase reporter activity. In addition, the SUnSET assay to measure newly synthesized cellular proteins showed that G2019S overexpression had a minimal effect on the total protein amount. However, we confirmed the previous result that G2019S overexpression increased the amount of protein synthesized from an exogenous gene, Flag-VAMP2, which was transfected as a reporter, whereas there was no significant change in the amount of the Flag-VAMP2 mRNA. Inhibition of protein degradation showed that protein accumulation in the vector control was higher than that of the G2019S overexpression vector. Our results suggest that LRRK2 protein influences the amount of protein by inhibiting protein degradation rather than by directly stimulating translation.

Astrocytes in injury states rapidly produce anti-inflammatory factors and attenuate microglial inflammatory responses

Kim, Jong-hyeon,Min, Kyoung-Jin,Seol, Wongi,Jou, Ilo,Joe, Eun-hye Blackwell Publishing Ltd 2010 Journal of Neurochemistry Vol.115 No.5

<P><I>J. Neurochem.</I> (2010) <B>115</B>, 1161–1171.</P><P>Abstract</P><P>Microglia are known to be a primary inflammatory cell type in the brain. However, microglial inflammatory responses are attenuated in the injured brain compared to those in cultured pure microglia. In the present study, we found that astrocytes challenged by oxygen–glucose deprivation (OGD) or H<SUB>2</SUB>O<SUB>2</SUB> released soluble factor(s) and attenuated microglial inflammatory responses. Conditioned medium prepared from astrocytes treated with OGD (OGD-ACM) or H<SUB>2</SUB>O<SUB>2</SUB> (H<SUB>2</SUB>O<SUB>2</SUB>-ACM) significantly reduced the levels of interferon-&ggr; (IFN-&ggr;)-induced microglial inflammatory mediators, including inducible nitric oxide synthase, at both the mRNA and protein levels. The anti-inflammatory effect of astrocytes appeared very rapidly (within 5 min), but was not closely correlated with the extent of astrocyte damage. Both OGD-ACM and H<SUB>2</SUB>O<SUB>2</SUB>-ACM inhibited STAT nuclear signaling, as evidenced by a reduction in both STAT-1/3 binding to the IFN-&ggr;-activated site and IFN-&ggr;-activated site promoter activity. However, both phosphorylation and nuclear translocation of STAT-1/3 was unchanged in IFN-&ggr;-treated microglia. The active component(s) in OGD-ACM were smaller than 3 kDa, and displayed anti-inflammatory effects independent of protein synthesis. These results suggest that, in the injured brain, astrocytes may act as a controller to rapidly suppress microglial activation.</P>

Oxidized DJ-1 Levels in Urine Samples as a Putative Biomarker for Parkinson's Disease

Jang, Jihoon,Jeong, Soyeon,Lee, Sung Ik,Seol, Wongi,Seo, Hyemyung,Son, Ilhong,Ho, Dong Hwan Hindawi 2018 Parkinson’s disease Vol.2018 No.-

<P>Parkinson's disease (PD) is the second most common neurodegenerative disease. Oxidative stress is the most critical risk factor for neurodegenerative diseases, including Alzheimer's disease (AD) and Huntington's disease (HD). Numerous reports have demonstrated that oxidative stress aggravates cytotoxicity in dopaminergic neurons and accelerates the formation of protein inclusions. In addition, oxidative stress, such as 4-hydroxynonenal (HNE), oxidized protein, and dopamine quinone, are related to PD progression. <I>DJ-1</I> is a PD-causative gene, and it plays a pivotal role as a sensor and eliminator of oxidative stress. Several studies have shown that oxidized DJ-1 (OxiDJ-1) formation is induced by oxidative stress. Hence, previous studies suggest that oxidized DJ-1 could be a biomarker for PD. We previously reported higher DJ-1 levels in Korean male PD patient urine exosomes than male non-PD controls. We speculate that OxiDJ-1 levels in PD patient urine might be higher than that in non-PD controls. In this study, we established an ELISA for OxiDJ-1 using recombinant DJ-1 treated with H<SUB>2</SUB>O<SUB>2</SUB>. Using Western blot assay and ELISA, we confirmed an increase of OxiDJ-1 from HEK293T cells treated with H<SUB>2</SUB>O<SUB>2</SUB>. Using our ELISA, we observed significantly higher, 2-fold, OxiDJ-1 levels in the urine of Korean PD patients than in non-PD controls.</P>

Increase in anti-apoptotic molecules, nucleolin, and heat shock protein 70, against upregulated LRRK2 kinase activity

Jang, Jihoon,Oh, Hakjin,Nam, Daleum,Seol, Wongi,Seo, Mi Kyoung,Park, Sung Woo,Kim, Hyung Gun,Seo, Hyemyung,Son, Ilhong,Ho, Dong Hwan ZOOLOGICAL SOCIETY OF KOREA 2018 ANIMAL CELLS AND SYSTEMS Vol.22 No.5

<P><B>ABSTRACT</B></P><P>Leucine-rich repeat kinase 2 (LRRK2) is involved in Parkinson’s disease (PD) pathology. A previous study showed that rotenone treatment induced apoptosis, mitochondrial damage, and nucleolar disruption via up-regulated LRRK2 kinase activity, and these effects were rescued by an LRRK2 kinase inhibitor. Heat-shock protein 70 (Hsp70) is an anti-oxidative stress chaperone, and overexpression of Hsp70 enhanced tolerance to rotenone. Nucleolin (NCL) is a component of the nucleolus; overexpression of NCL reduced cellular vulnerability to rotenone. Thus, we hypothesized that rotenone-induced LRRK2 activity would promote changes in neuronal Hsp70 and NCL expressions. Moreover, LRRK2 G2019S, the most prevalent LRRK2 pathogenic mutant with increased kinase activity, could induce changes in Hsp70 and NCL expression. Rotenone treatment of differentiated SH-SY5Y (dSY5Y) cells increased LRKK2 levels and kinase activity, including phospho-S935-LRRK2, phospho-S1292-LRRK2, and the phospho-moesin/moesin ratio, in a dose-dependent manner. Neuronal toxicity and the elevation of cleaved poly (ADP-ribose) polymerase, NCL, and Hsp70 were increased by rotenone. To validate the induction of NCL and Hsp70 expression in response to rotenone, cycloheximide (CHX), a protein synthesis blocker, was administered with rotenone. Post-rotenone increased NCL and Hsp70 expression was repressed by CHX; whereas, rotenone-induced kinase activity and apoptotic toxicity remained unchanged. Transient expression of G2019S in dSY5Y increased the NCL and Hsp70 levels, while administration of a kinase inhibitor diminished these changes. Similar results were observed in rat primary neurons after rotenone treatment or G2019S transfection. Brains from G2019S-transgenic mice also showed increased NCL and Hsp70 levels. Accordingly, LRRK2 kinase inhibition might prevent oxidative stress-mediated PD progression.</P><P><B>Abbreviations:</B> 6-OHDA: 6-hydroxydopamine; CHX: cycloheximide; dSY5Y: differentiated SH-SY5Y; g2019S tg: g2019S transgenic mouse; GSK/A-KI: GSK2578215A kinase inhibitor; HSP70: heat shock protein 70; LDH: lactose dehydrogenase; LRRK2: leucine rich-repeat kinase 2; MPTP: 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine; myc-GS LRRK2: myc-tagged g2019S LRRK2; NCL: nucleolin; PARP: poly(ADP-ribose) polymerase; PD: Parkinson’s disease; PINK1: PTEN-induced putative kinase 1; pmoesin: phosphorylated moesin at t558; ROS: reactive oxygen species</P>

Estrogen Receptor-alpha Mediates the Effects of Estradiol on Telomerase Activity in Human Mesenchymal Stem Cells.

Cha, Young,Kwon, Su Jin,Seol, Wongi,Park, Kyung-Soon Korean Society for Molecular Biology 2008 Molecules and cells Vol.26 No.5

<P>Sex steroid hormone receptors play a central role in modulating telomerase activity, especially in cancer cells. However, information on the regulation of steroid hormone receptors and their distinct functions on telomerase activity within the mesenchymal stem cell are largely unavailable due to low telomerase activity in the cell. In this study, the effects of estrogen (E2) treatment and function of estrogen receptor alpha (ERalpah) and estrogen receptor beta (ERbeta) on telomerase activity were investigated in human mesenchymal stem cells (hMSCs). Telomerase activity and mRNA expression of the catalytic subunit of telomerase (hTERT) were upregulated by treatment of the cells with E2. The protein concentration of ER alpah was also increased by E2 treatment, and enhancement of ER alpah accumulation in the nucleus was clearly detected with immunocytochemistry. When ER alpah expression was reduced by siRNA transfection into hMSCs, the effect of E2 on the induction of hTERT expression and telomerase activity was diminished. In contrast, the transient overexpression of ER alpah increased the effect of E2 on the expression of hTERT mRNA. These findings indicate that the activation of hTERT expression and telomerase activity by E2 in hMSCs depends on ER alpah, but not on ERbeta.</P>

Hypoxia-inducible factor-1α upregulates tyrosine hydroxylase and dopamine transporter by nuclear receptor ERR&ggr; in SH-SY5Y cells

Lim, Juhee,Kim, Hyo-In,Bang, Yeojin,Seol, Wongi,Choi, Hueng-Sik,Choi, Hyun Jin Wolters Kluwer Health | Lippincott Williams Wilkin 2015 NEUROREPORT - Vol.26 No.6

Hypoxia-inducible factor-1α (HIF-1α) is a transcription factor relevant to the development of many mammalian organs including the brain. However, the molecular mechanisms by which signaling events mediate neuronal differentiation have not been fully elucidated. In the present study, we show for the first time that the orphan nuclear receptor estrogen-related receptor &ggr; (ERR&ggr;) is upregulated by HIF-1α and plays essential roles in HIF-1α-induced upregulation of dopaminergic marker molecules such as tyrosine hydroxylase and dopamine transporter. We found that deferoxamine upregulated HIF-1α and enhanced the dopaminergic phenotype and neurite outgrowth of SH-SY5Y cells. Deferoxamine activated transcription and protein expression of ERR&ggr;, and deferoxamine-induced upregulation of tyrosine hydroxylase and dopamine transporter was attenuated by using the ERR&ggr; inverse agonist or silencing ERR&ggr;. Altogether, these results suggest that HIF-1α can positively regulate the dopaminergic phenotype through ERR&ggr;. This study could provide new perspectives for understanding the mechanisms underlying the promotion of dopaminergic neuronal differentiation by hypoxia.

Estrogen Receptor-α Mediates the Effects of Estradiol on Telomerase Activity in Human Mesenchymal Stem Cells

Young Cha,Su Jin Kwon,Wongi Seol,박경순 한국분자세포생물학회 2008 Molecules and cells Vol.26 No.5

Sex steroid hormone receptors play a central role in modulating telomerase activity, especially in cancer cells. However, information on the regulation of steroid hormone receptors and their distinct functions on telomerase activity within the mesenchymal stem cell are largely unavailable due to low telomerase activity in the cell. In this study, the effects of estrogen (E₂) treatment and function of estrogen receptor alpha (ER α) and estrogen receptor beta (ER β) on telomerase activity were investigated in human mesenchymal stem cells (hMSCs). Telomerase activity and mRNA expression of the catalytic subunit of telomerase (hTERT) were upregulated by treatment of the cells with E2. The protein concentration of ER alpha was also increased by E₂ treatment, and enhancement of ER alpha accumulation in the nucleus was clearly detected with immunocytochemistry. When ER alpha expression was reduced by siRNA transfection into hMSCs, the effect of E₂ on the induction of hTERT expression and telomerase activity was diminished. In contrast, the transient overexpression of ERα increased the effect of E₂ on the expression of hTERT mRNA. These findings indicate that the activation of hTERT expression and telomerase activity by E₂ in hMSCs depends on ER alpha, but not on ERβ