The Existence of a Putative Regulatory Element in 3'-Untranslated Region of Proto-oncogene HOX11's mRNA

Li, Yue,Jiang, Zhao-Zhao,Chen, Hai-Xu,Leung, Wai-Keung,Sung, Joseph J.Y.,Ma, Wei-Jun Korean Society for Biochemistry and Molecular Biol 2005 Journal of biochemistry and molecular biology Vol.38 No.4

HOX11 encodes a homeodomain-containing transcription factor which directs the development of the spleen during embryogenesis. While HOX11 expression is normally silenced through an unknown mechanism in all tissues by adulthood, the deregulation of HOX11 expression is associated with leukemia, such as T-cell acute lymphoblastic leukemia. The elucidation of regulatory elements contributing to the molecular mechanism underlying the regulation of HOX11 gene expression is of great importance. Previous reports of HOX11 regulatory elements mainly focused on the 5'-flanking region of HOX11 on the chromosome related to transcriptional control. To expand the search of putative cis-elements involved in HOX11 regulation at the post-transcriptional level, we analyzed HOX11 mRNA 3'-untranslated region (3'UTR) and found an AU-rich region. To characterize this AU-rich region, in vitro analysis of HOX11 mRNA 3'UTR was performed with human RNA-binding protein HuR, which interacts with AU-rich element (ARE) existing in the 3'UTR of many growth factors' and cytokines' mRNAs. Our results showed that the HOX11 mRNA 3'UTR can specifically bind with human HuR protein in vitro. This specific binding could be competed effectively by typical ARE containing RNA. After the deletion of the AU-rich region present in the HOX11 mRNA 3'UTR, the interaction of HOX11 mRNA 3'UTR with HuR protein was abolished. These findings suggest that HOX11 mRNA 3'UTR contains cis-acting element which shares similarity in the action pattern with RE-HuR interactions and may involve in the post-transcriptional regulation of the HOX11 gene.

흰쥐에서 임신 중 레티노산이 입천장 형태발생에 미치는 영향

정명섭(Meang Sub Cheng),유병기(Byoung Ki Yoo),박형우(Hyoung Woo Park),김명희(Myoung Hee Kim) 대한해부학회 2006 Anatomy & Cell Biology Vol.39 No.4

흰쥐 입천장발생은 배자 발생 14일에서 17일까지 입천장선반 (palatal shelves)이 아래쪽으로 성장하고 16일에 혀 위쪽으로 이동하여 중앙을 중심으로 병합하여 17일에는 완전한 입천장이 형성된다. 레티노산은 비타민 A의 생체 내 활성 대사산물로써 발생에서 배자의 앞뒤축 형성 등에 관여하는데, 이런 레티노산에 과다히 노출∙결핍 시에는 기형이 유발되고, 배자 축 형성에 관여하는 Hox 유전자의 발현 또한 조절한다고 알려져 있다. 본 연구는 임신중 레티노산이 입천장 형태발생에 미치는 영향을 보기위하여 임신 11일째 흰쥐에 레티노산을 복강내주사로 주입하고 발생 13일부터 17일까지의 배자를 분리한 후 배자의 형태를 분석하고 또 형태형성에 관여하는 Hox와 Bcl-2 family 유전자를 포함하여 레티노산에 의해 차등 발현하는 유전자를 (DD) RT-PCR방법을 이용하여 분석하였다. 실험 결과 레티노산에 노출된 실험군 배자의 경우 정상군에 비해 머리 발생 지연을 포함하여 머리-엉덩 길이와 사지 등의 길이가 짧았다. 또 발생 17일째에는 정상군에서는 완전한 입천장이 형성되었으나 실험군에서는 총 52예의 배자 중 36예에서 입천장갈림증이 관찰되었으며, 입천장갈림증 간격은 약 0.80±0.36 mm이었다. DDRT-PCR 결과 레티노산 투여로 인해 차등발현하는 유전자들이 많이 존재함이 관찰되었다. Hoxa7 유전자는 정상군에서는 발생 13일째에 그러나 레티노산 투여군에서는 발생 15일째에 강하게 발현한 후 점차 감소한 반면 Hoxc8은 발현이 거의 감지되지 않았으나 정상군에서는 발생 16일째에서 그리고 레티노산 투여군에서는 15일째에서 아주 약한 발현이 감지되었다. Bcl-2 family 유전자의 경우 정상군에서는 발생 13일에서 17일까지 Bclxl과 Bcl-2 유전자, 그리고 Bax 유전자 모두 강하게 발현하였으나, 실험군의 경우 Bcl-xl과 Bcl-2의 발현이 발생 16일 이후에는 감소한 반면 Bax의 경우는 발생 13일에서 15일까지 매우 저하되었다. 이상의 결과로 임신중 레티노산 노출이 배자 발생과정 중 입천장갈림증을 포함한 형태적 기형유발을 가능하게 하는 물질임을 알 수 있었으며, 또 배자의 형태 발생에 관련된 Hox 유전자와 세포사멸 관련 유전자인 Bcl-xl, Bcl-2, 그리고 Bax 유전자의 발현을 조절하여 배자, 특히 입천장의 형태 기형 유발에 관여한 것으로 유추된다. In order to understand the effect of retinoic acid (RA) on the craniofacial pattern formation during embryogenesis, we injected RA intraperitoneally into the pregnant female rat on day 11 post coitum (p.c.) and then embryos of day 13 to day 17 p.c. were isolated consequently. The overall morphology and the differential gene expression patterns were analyzed by the microscopic and (DD) RT-PCR methods, respectively. For the morphological study, the retardation of craniofacial region, the shortage of crown rump length and limbs were analyzed in the RA-treated embryos. In the RA-treated embryos of day 17, it was observed that the palatogenesis was completely finished just like in the normal embryos. However, the cleft plate was observed in 36 out of 52 total samples with the distance of cleft palate being 0.80±0.36 mm in average. The temporal expression pattern of Hox genes through RT-PCR revealed that the expression of Hoxa7 reached its peak on day 13 then slowly declined in the normal embryos. Whereas in the RA-treated embryos, the expression peak was observed on day 15, then declined subsequently. With the Hoxc8 gene, its expression was low in all stages until the day 16 of normal embryogenesis. On the other hand, Hoxc8 gene expression was detected slightly early on day 15 in the RA-treated embryos. In the study of Bcl-2 family genes, uniformly strong expression of anti-apoptotic and pro-apoptotic genes was observed from day 13 to day 17 of normal embryos, whereas anti-apoptotic gene expressions were decreased after day 16 in the RAtreated embryos. Additionally, a dramatic decline of pro-apoptotic gene expression was observed from day 13 to day 15 of the RA-treated embryos. Therefore, we believe that RA is a potential factor that is actively involved in the cleft palate formation. Moreover, it is profoundly linked with the regulation of Hox and Bcl-2 family gene expression pattern that leads to the embryonic malformation.

A Study of Hox Gene Expression Profile During Murine Liver Regeneration

Youn, Boyeon,Kim, Byung-Gyu,Kim, Myoung Hee 대한의생명과학회 2003 Biomedical Science Letters Vol.9 No.1

Liver is an organ having an ability to regenerate by itself when it is damaged or removed. Since the research on the liver regeneration so far was regarding on the cellular multiplications not the formation of the shape, we intended to analyze the expression pattern of Hox genes during liver regeneration. RNA samples isolated from liver at the time of partial hepatectomy, 4 hours as well as 3 days later following regeneration were used to perform RT-PCR with Hox-specific degenerate primers. The PCR products were cloned, sequenced and analyzed through BLAST program, Genes belonging to the AbdB type Hox genes (paralogous groups IX-XIII)expressed predominantly during regeneration, while the other group (I-VII), especially Hoxal and b1 seemed to be expressed continuously before and after regeneration. These data altogether imply that paralogous group IX and X genes including Hoxa10 and d10 seemed to be regeneration specific genes of liver.

A Phylogenetic Analysis for Hox Linked Gene Families of Vertebrates

김선우,정길라,이재현,박하영,김창배 한국통합생물학회 2008 Animal cells and systems Vol.12 No.4

The human chromosomes 2, 7, 12 and 17 show genomic homology around Hox gene clusters, is taken as evidence that these paralogous gene families might have arisen from a ancestral chromosomal segment through genome duplication events. We have examined protein data from vertebrate and invertebrate genomes to analyze the phylogenetic history of multi-gene families with three or more of their representatives linked to human Hox clusters. Topology comparison based upon statistical significance and information of chromosome location for these genes examined have revealed many of linked genes coduplicated with Hox gene clusters. Most linked genes to Hox clusters share the same evolutionary history and are duplicated in concert with each other. We conclude that gene families linked to Hox clusters may be suggestion of ancient genome duplications. The human chromosomes 2, 7, 12 and 17 show genomic homology around Hox gene clusters, is taken as evidence that these paralogous gene families might have arisen from a ancestral chromosomal segment through genome duplication events. We have examined protein data from vertebrate and invertebrate genomes to analyze the phylogenetic history of multi-gene families with three or more of their representatives linked to human Hox clusters. Topology comparison based upon statistical significance and information of chromosome location for these genes examined have revealed many of linked genes coduplicated with Hox gene clusters. Most linked genes to Hox clusters share the same evolutionary history and are duplicated in concert with each other. We conclude that gene families linked to Hox clusters may be suggestion of ancient genome duplications.

Differential Expression of Hox Genes during Murine Liver Regeneration

Boyeon Youn,Hyoung Woo Park,Sung Goo Park,Myoung Hee Kim 한국유전학회 2007 Genes & Genomics Vol.29 No.4

In order to infer the Hox code during liver regeneration, Hox gene expression pattern was analyzed during liver regeneration after partial hepatectomy in mice. Total RNAs and proteins have been prepared from the liver tissues at various times after regeneration (0 h, 4 h, and 3 d) following partial hepatectomy. Using degenerate RT-PCR, cloning, and sequencing technique, expressed Hox genes were analyzed. The Hox genes belonging to the paralogous groups VIII, IX and X manifested themselves in a peculiar way during regeneration process with stage specific characteristics: Hox genes belonging to the paralogous groups IX and X were almost exclusively expressed during regeneration whereas the paralogous group VIII, especially Hoxb8 and -c8 were strongly expressed in normal liver. The proteom analysis revealed that almost 21% of the differentially expressed proteins (6 out of 29 differentially expressed protein spots) during liver regeneration turned out to be Hox genes. Furthermore, Hoxa10 and- d10 belonging to the AbdB type paralogous group X were upregulated during regeneration. These results altogether indicate that Hox genes seemed to be differentially expressed during murine liver regeneration at the transcriptional as well as translational level. Especially the Hox genes belonging to the paralogous group X such as Hoxa10 and Hoxd10 seemed to be a part of Hox code for the murine liver regeneration.

피부종양에서 HOX D3와 혈관성장인자(VEGF-C)의 발현

김형동 ( Hyung Dong Kim ),조문균 ( Moon Kyun Cho ),박영립 ( Young Lip Park ),이종석 ( Jong Suk Lee ),황규왕 ( Kyu Uang Whang ) 대한피부과학회 2007 大韓皮膚科學會誌 Vol.45 No.4

Background: The anatomical relation between a malignant tumor and its vascular and lymphatic bed is an important influencing metastasis. Hox D3 is required for these expressions of integrin αvβ3 and urokinase plasminogen activator (uPA), which contribute to endothelial cell adhesion, invasion, and migration during angiogenesis. Recent studies in different tumor types have shown that vascular endothelial growth factor-C (VEGF-C), which displays a high specificity for lymphatic endothelium, is involved in tumor-induced lymphagiogenesis and lymphatic metastatic spread. Objective: This study was designed to measure the expression of HOX D3 and VEGF-C in different skin cancers. Methods: The expression of HOX D3 and VEGF-C was examined by immunohistochemical staining of 40 skin cancer tissue samples, including 8 keratoacanthomas, 8 extramammary paget`s disease, 8 basal cell carcinomas, 8 squamous cell carcinomas and 8 malignant melanomas. Results: Immunohistochemical analysis of 40 skin cancer tissue samples revealed a high expression of HOX D3 and VEGF-C in the more aggressive and invasive skin tumors, including squamous cell carcinomas and malignant melanomas. On the other hand, low expression was seen in the less-invasive skin tumors, including keratoacanthomas, extramammary paget`s disease and basal cell carcinomas. Also the degree of expression of HOX D3 and VEGF-C showed a statistically-significant correlation with each skin tumor (p<0.05). Conclusion: These findings provide evidence that the upregulation of HOX D3 and VEGF-C might be involved in the promotion of angiogenesis and lymphagiogenesis in skin tumors and play an important role in metastasis. (Korean J Dermatol 2007;45(4):354~361)

북방전복 (Haliotis discus hannai) 의 Hox 유전자 동정 및 발현 분석

박은희,신은하,노은수,김영옥,박중연,남보혜 한국패류학회 2019 The Korean Journal of Malacology Vol.35 No.1

Hox genes are expressed along the anterior-posterior body axis in a collinear fashion in the majority of bilaterians. Contrary to polyplacophorans, gastropods including abalone deviate from this pattern by expressing Hox genes in distinct morphological structures and not in a staggered fashion. However, the molecular developmental understandings remain unknown. In the present study, we isolated DNA sequence of Hox gene from genome database and investigated its mRNA expression in early developmental stages of abalone, Haliotis discus hannai. The H. discus hannai Hox gene (HDH-HoxA2) showed 36% and 37% amino acid identities with with HoxA2 of Crassostrea gigas and Mixuhopecten yessoensis, respectively. The 62 amino acids sequence of homeodomain was conserved well in HDH-HoxA2. The qRT-PCR detected HDH-HoxA2 at the first day after hatching (trocophore larvae stage), but the expression of HDH-HoxA2 mRNA decreased after two days post-hatching. The present study is the first to report on the molecular cloning, characterization and expression analysis of Hox gene in H. discus hannai. The results might provide new insights into the regulation of early developmental stages of abalone.

Effects of Dexamethasone on Embryo Development and Hox Gene Expression Patterns in Mice

Sang Hoon Kim,Ji-Yeon Lee,Sung Joo Park,Myoung Hee Kim 대한의생명과학회 2011 Biomedical Science Letters Vol.17 No.3

During pregnancy, stress induces maternal glucocorticoid secretion, which in turn is known to affect structural malformation, retardation of growth, reduced birth weight of the fetus. As Hox genes are master transcription factors which fulfill critical roles in embryonic development, we aimed to explore the possibility that alterations of the Hox gene expression might be involved in stress-induced malformation. The pregnant mice were injected with dexamethasone at a dose of 1 mg/kg or 10 mg/kg on day 7.5, 8.5 and 9.5 p.c. (post coitum), as well as saline as control. Embryos of E11.5 and E18.5 were obtained by sacrificing pregnant animals. Weight and crown-rump length (CRL) were measured. RT-PCR was performed to examine the Hox gene expression levels. Embryos given dexamethasone at day 7.5~9.5 p.c. had small CRL and weighed less both in E11.5 and E18.5. The percentage of embryos showing abnormalities was high in groups received high dose of dexamethasone. To define the molecular basis for abnormal embryonic development, we analyzed the Hox gene expression pattern and found that many Hox genes display altered expression. Effects of prenatal dexamethasone treatment on embryonic development might be associated with the aberrant Hox gene expression.

개불에서 Hox 유전자의 동정 및 계통분석

신길상,이대희 순천향대학교 기초과학연구소 2004 순천향자연과학연구 논문집 Vol.10 No.2

An echiuriod species, Urechis unicinctus, was surveyed for Homeobox(Hox) genes using PCR with Homeobox-specific degenerate primers. We were able to identify five Hox genes. The results revealed that the five Hox genes were belonged to central group of body plan-genes in the echiuroid. Analyses in this work made it able to classify the five Hox genes comparatively and phylogenetically, and came to the conclusion that the echiuroid can be classified one of lophotrochozoan. The result may be the first report regarding sequence information and phylogenetic analysis of Hox genes in the echiuroid species.

A Phylogenetic Analysis for Hox Linked Gene Families of Vertebrates

Kim, Sun-Woo,Jung, Gi-La,Lee, Jae-Hyoun,Park, Ha-Young,Kim, Chang-Bae The Korean Society for Integrative Biology 2008 Animal cells and systems Vol.12 No.4

The human chromosomes 2, 7, 12 and 17 show genomic homology around Hox gene clusters, is taken as evidence that these paralogous gene families might have arisen from a ancestral chromosomal segment through genome duplication events. We have examined protein data from vertebrate and invertebrate genomes to analyze the phylogenetic history of multi-gene families with three or more of their representatives linked to human Hox clusters. Topology comparison based upon statistical significance and information of chromosome location for these genes examined have revealed many of linked genes coduplicated with Hox gene clusters. Most linked genes to Hox clusters share the same evolutionary history and are duplicated in concert with each other. We conclude that gene families linked to Hox clusters may be suggestion of ancient genome duplications.