• 대한의생명과학회지
• /
• 제9권3호
• /
• pp.159-165
• /
• 2003

Ankyrins are a ubiquitously expressed family of intracellular adaptor proteins involved in targeting diverse proteins to specialized membrane domains in both the plasma membrane and the endoplasmic reticulum. Recently, the studies with C-terminus of ankyrins have identified that ankyrin-B is capable of interacting with Hsp40 and sAnkl is capable of interacting with obscurin and titin, but the function of C-terminal domain of ankyrin-G remains unknown. To identify proteins interacting C-terminus of ankyrin-G, we used the C-terminus of ankyrin-G as a bait for a yeast two-hybrid screen of brain cDNA library. Approximately 1.33$\times$l0$^6$ transformants were screened, of which 13 positive clones were obtained as determined by activation of HIS3, ADE2 and MELl reporter genes. Sequence analyses of these 13 plasmids revealed that cDNA inserts of 13 colonies showed highly homologous to 11 genes, including 5 known (i.e., Na$^+$/K$^+$ ATPase $\beta$1, SERBPl, UTF2, cytochrome C oxidase and collagen IV $\alpha$2) and 6 unknown genes. The evaluation of the proteins that emerge from these experiments provides a rational approach to investigate the those proteins significant in interaction with ankyrin-G.

• 대한의생명과학회지
• /
• 제9권2호
• /
• pp.105-110
• /
• 2003

Ankyrins are a ubiquitously expressed family of intracellular adaptor proteins involved in targeting diverse proteins to specialized membrane domains in both the plasma membrane and the endoplasmic reticulum. Canonical ankyrins are 190-220 kDa proteins expressed in most tissues and cell types and comprise a membrane-binding domain (MBD) of 24 ANK repeats, a spectrin-binding domain, a death domain and a C-terminal domain. Rescue studies with ankyrin-B/G chimeras have identified the C-terminal domain of ankyrin-B as the defining domain in specifying ankyrin-B activity, but the function of C-terminal domain of ankyrin-B is, however, not known. We report here that the C-terminal domain of ankyrin-B is capable of interacting with the C-terminal Region of Hsp40. The Hsps are induced not only by heat shock but also by various other environmental stresses. Hsps are also expressed constitutively at normal growth temperatures and have basic and indispensable functions in the life cycle of proteins as molecular chaperones, as well as playing a role in protecting cells from the deleterious stresses. The binding sites required in the interaction between C-terminal domain of ankyrin-B and C-terminal region of Hsp40 were characterized using the yeast two-hybrid system and GST-pull down assay. The interaction between ankyrin-B and Hsp40 represents the first direct evidence of ankyrin's role as chaperones.

• Molecules and Cells
• /
• 제37권12호
• /
• pp.881-887
• /
• 2014

Cell proliferation is tightly controlled by the cell-cycle regulatory proteins, primarily by cyclins and cyclin-dependent kinases (CDKs) in the $G_1$ phase. The ankyrin repeat-rich membrane spanning (ARMS) scaffold protein, also known as kinase D-interacting substrate of 220 kDa (Kidins 220), has been previously identified as a prominent downstream target of neurotrophin and ephrin receptors. Many studies have reported that ARMS/Kidins220 acts as a major signaling platform in organizing the signaling complex to regulate various cellular responses in the nervous and vascular systems. However, the role of ARMS/Kidins220 in cell proliferation and cell-cycle progression has never been investigated. Here we report that knockdown of ARMS/Kidins220 inhibits mouse neuroblastoma cell proliferation by inducing slowdown of cell cycle in the $G_1$ phase. This effect is mediated by the upregulation of a CDK inhibitor p21, which causes the decrease in cyclin D1 and CDK4 protein levels and subsequent reduction of pRb hyperphosphorylation. Our results suggest a new role of ARMS/Kidins220 as a signaling platform to regulate tumor cell proliferation in response to the extracellular stimuli.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제37권2호
• /
• pp.49-54
• /
• 2018

To artificially enhance antimicrobial peptide expression in Bombyx mori, we constructed genetically engineered silkworms overexpressing Rel family transcription factor. The truncated BmRelish1 (BmRelish1t) gene contained a Rel homolog domain (RHD), nuclear localization signal (NLS), acidic and hydrophobic amino acid (AHAA)-rich region, and death domain (DD), but no ankyrin-repeat (ANK) domain. The BmRelish1t gene was controlled by B. mori cytoplasmic actin 3 promoter in the PiggyBac transposon vector. Chromosome analysis of G1 generations of a transgenic silkworm with EGFP expression confirmed stable insertion of BmRelish1t. BmRelish1t gene overexpression in transgenic silkworms resulted in higher mRNA expression levels of B. mori antimicrobial peptides such as lebocin(~20.5-fold), moricin(~8.7-fold), and nuecin(~17.4-fold) than those in normal silkworms.

• BMB Reports
• /
• 제42권9호
• /
• pp.568-573
• /
• 2009

Fatty acid oxidation (FAO) defects cause abnormal lipid accumulation in various tissues, which provides an opportunity to uncover novel genes that are involved in lipid metabolism. During a gene expression study in the riboflavin deficient induced FAO disorder in the chicken, we discovered the dramatic increase in mRNA levels of an uncharacterized gene, ANKRD9. No functions have been ascribed to ANKRD9 and its orthologs, although their sequences are well conserved among vertebrates. To provide insight into the function of ANKRD9, the expression of ANKRD9 mRNA in lipidperturbed paradigms was examined. The hepatic mRNA level of ANKRD9 was repressed by thyroid hormone ($T_3$) and fasting, elevated by re-feeding upon fasting. However, ANKRD9 mRNA level is reduced in response to apoptosis. Transient transfection assay with green fluorescent protein tagged- ANKRD9 showed that this protein is localized within the cytoplasm. These findings point to the possibility that ANKRD9 is involved in intracellular lipid accumulation.

• Asian-Australasian Journal of Animal Sciences
• /
• 제18권4호
• /
• pp.483-489
• /
• 2005

To understand the molecular mechanisms that regulate intramuscular fat deposition and its release, cDNA clones expressed in adipose tissues of Korean cattle were identified by differential screening from adipose tissue cDNA library. By partial nucleotide sequencing of 486 clones and a search for sequence similarity in NCBI nucleotide databases, 245 clones revealed unique clones. By a functional grouping of the clones, 14% of the clones were categorized to metabolism and enzyme-related group (stearoyl CoA desaturase, lactate dehydrogenase, fatty acid synthase, ATP citrate lyase, lipoprotein lipase, acetyl CoA synthetase, etc), and 6% to signal transduction/cell cycle-related group (C/EBP, cAMP-regulated phosphoprotein, calmodulin, cyclin G1, cyclin H, etc), and 4% to cytoskeleton and extracellular matrix components (vimentin, ankyrin 2, gelosin, syntenin, talin, prefoldin 5). The obtained 245 clones will be useful to study lipid metabolism and signal transduction pathway in adipose tissues and to study obesity in human. Some clones were subjected to full-sequencing containing open reading frame. The cDNA clone of bovine homolog of human prefoldin 5 gene had a total length of 959 nucleotides coding for 139 amino acids. Comparison of the deduced amino acid sequences of bovine prefoldin 5 with those of human and mouse showed over 95% identity. The cDNA clone of bovine homolog of human ubiquitin-like/S30 ribosomal fusion protein gene had a total length of 484 nucleotides coding for 133 amino acids. Comparison of the deduced amino acid sequences of bovine ubiquitin-like/S30 ribosomal fusion protein gene with those of human, rat and mouse showed over 97% identity. The cDNA clone of bovine homolog of human proteolipid protein 2 mRNA had a total length of 928 nucleotides coding for 152 amino acids. Comparison of the deduced amino acid sequences of bovine proteolipid protein 2 with those of human and mouse showed 87.5% similarity. The cDNA clone of bovine homolog of rat thymosin beta 4 had a total length of 602 nucleotides coding for 44 amino acids. Comparison of the deduced amino acid sequences of bovine thymosin beta 4 gene with those of human, mouse and rat showed 93.1% similarity. The cDNA clone of bovine homolog of human myotrophin mRNA had a total length of 790 nucleotides coding for 118 amino acids. Comparison of the deduced amino acid sequences of bovine myotrophin gene with those of human, mouse and rat showed 83.9% similarity. The functional role of these clones in adipose tissues needs to be established.

• 한국치위생학회지
• /
• 제24권3호
• /
• pp.219-227
• /
• 2024

연구목적: 치수는 신경과 혈관을 포함하고 있는 부위로 다양한 자극이나 세균에 의해 염증이 생기면 이를 치수염이라고 한다. 치수염은 말초 신경조직 변화와 심한 통증을 유발하는 질환으로 만성적 통증을 유발하나, 삼차신경절의 신경세포 활성화와 특정 나트륨 채널(Nav1.7) 발현 사이의 관계는 잘 알려져 있지 않다. 이에 본 연구에서는 실험적으로 유도된 치수염이 말초신경에서 Nav1.7 나트륨 채널의 발현을 활성화시켜, 삼차신경절의 뉴런을 활성화함으로써 통증을 유발한다는 사실을 발견하고 이를 조절하는 신호기전을 규명하고자 하였다. 연구방법: 실험동물(Male C57BL/6 생쥐, 6주, 20-25 g)의 상악 제1대구치 치수에 AITC를 처리하여 급성 치수염을 유발하고, 3일 후 실시간 광영상 이미지를 이용하여 삼차신경절의 뉴런 활성화를 측정 및 비교 분석하였다. 단백질 분석을 통해 뇌간(SpVc)에서 치수염 통증유발 신호조절기전에 관여하는 여러가지 단백질들(p-ERK, c-FOS, TRPA1, p-CRMP2)의 발현을 관찰하였다. 연구결과: 시공간적 광영상 이미지를 통해 삼차신경절의 뉴런세포들은 대조군과 비교 시 급성 치수염 모델에서 흥분성 활성화가 유도되어 신경학적 변화가 일어남을 관찰하였다. 또한 조직학 및 분자생물학적 결과를 통해 치수염으로 인한 특정 나트륨 채널(Nav1.7)의 증가가 통증을 유발한다는 사실을 확인하였다. 또한, ProTxII(Nav1.7의 선택적 억제 약물) 처리를 통해 뉴런의 과흥분성 활성이 억제됨에 따라, 치수염이 삼차신경절에서 나트륨 채널(Nav1.7)의 증가를 유도하고 이러한 특정 나트륨 채널을 효과적으로 제어하면 치수염의 통증을 줄이는 방법이 될 것이라 생각된다. 결론: 본 연구의 결과를 통해 과 발현된 특정 나트륨 채널(Nav1.7)을 억제하면 삼차신경절에서 통각 신호 처리를 조절하고 치수염에 의해 유발되는 통증을 효과적으로 조절할 수 있음을 시사한다.

• 한국국제농업개발학회지
• /
• 제23권5호
• /
• pp.537-545
• /
• 2011

본 연구는 간척지에서 재배가 가능한 내염성 보리 품종육성을 위한 기초정보를 얻고자 겉보리 두 품종을 대상으로 생육 초기 염 스트레스에 따른 생리적 반응과 잎 프로테옴의 발현 양상 변화를 분석한 결과는 아래와 같다. 1. 토양의 염 농도가 증가함에 따라 보리의 건물중은 무처리구에 비해 유의적으로 감소하는 경향이었으며, 상록보리는 무처리구에 비해 건물중 감소가 작았으며, 선우보리는 컸다. 2. 염처리에 따른 잎의 엽록소 함량을 나타내는 SPAD 값은 상록보리가 57.6으로 47.6인 선우보리보다 높았으며, Na⁺의 함량은 선우보리에서 유의적으로 높았고, K⁺/Na⁺의 비율은 상록 보리에서 높은 경향을 보였다. 3. 이차원전기영동에 의하여 염 스트레스에 의한 잎 프로테옴의 발현양상을 분석한 결과 47개 단백질 spot이 발현양의 차이를 나타냈다. 품종별로 발현양이 증가한 단백질 spot은 상록보리와 선우보리에서 각각 17개와 14개로 나타났고, 발현양이 감소한 단백질 spot은 상록보리와 선우보리에서 각각 28개 및 27개로 확인되었다. 4. 염처리에 따른 발현양의 차이를 보이는 18개 단백질을 동정한 결과 ribosomal protein 등 기능과 스트레스와의 관련성이 보고된 10개의 단백질과 ankyrin repeat domain protein 등 스트레스 조건에서의 역할이 명확하지 않은 4개의 단백질 및 Os02g0753300 등 기능 및 스트레스와의 관련성을 알 수 없는 2개의 단백질이 동정되었다.