말에서 dynein, cytoplasmic 1, light intermediate chain 2(DYNC1LI2) alternative transcripts의 분자생물학적 분석
Molecular Analysis of Alternative Transcripts of the Equine Dynein, Cytoplasmic 1, Light Intermediate Chain 2(DYNC1LI2) Gene
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말에서 주요 경제형질인 운동과 관련된 연구는 중요하지만, 현재까지의 연구는 물리학적, 생리학적연구에 치중되어 있어 분자수준의 연구는 미비한 실정이다. 이에 본 연구는 선행연구를 통하여 경주마에서 RNA-sequencing을 수행하여 운동 전·후 alternative transcript 이형에 따라 발현 양상이 상이한 유전자(DYNC1LI2, COBLL1, AXL, PLEKHG1)를 발굴하였다. 이 중, DYNC1LI2 유전자를 선택하여분자생물학적 분석 및 운동성과의 관계에 대하여 연구를 수행하였다. 그 결과, DYNC1LI2 유전자의 2가지 전사 이형은 긴 형태의 전사체(DYNC1LI2a)와 결손이 일어나 상대적으로 짧아진 전사체(DYNC1LI2b)의 형태로 존재하는 것을 확인하였고, 두 가지 전사 이형 모두가 말의 각 조직(갑상선, 결장, 골격근, 맹장, 심장, 신장, 척수, 폐)에 존재함을 확인하였다. 또한, 운동 전과 운동 후 발현량 분석을 통해 두 가지 전사 이형이 동일하게 운동에 따라 발현이 감소하는 것을 확인하였다. 추가적으로 두가지 전사 이형의 아미노산 비교 분석 결과, 엑손영역에 결손이 일어나는 부분은 단백질의 인산화 및당질화와 관련이 있음을 확인하였다. 이는 DYNC1LI2a가 DYNC1LI2b에 비해 더욱 단백질의 안정화 작용을 하는 것을 의미하며, DYNC1LI2 유전자가 운동에 따라 발현이 달라짐에 따라 차후 말에서 운동관련 연구에 대한 기반 자료로써 사용될 수 있음을 시사한다.
Abstract
General economy trait related exercise of horse is important, but researches which is studieduntil now is mostly about physical and physiological research related with horse athletic ability,researches in molecular level hasn't been studied enough. In previous study, RNA-sequencingwas conducted with skeletal muscle and blood in 6 thoroughbreds, and through RNA-sequencing,four genes(DYNC1LI2, COBLL1, AXL, PLEKHG1) which is expressed differently according toAlternative splicing isoform before and after exercise are predicted as a result. Among thosegenes, we validated DYNC1LI2 gene and investigated for relationship between DYNC1LI2 gene and athletic ability. Our research team validated the presence of alternative spliced isoforms ofequine DYNC1LI2, namely DYNC1LI2a as a long form and DYNC1LI2b as a short form invitro. Also as a result for RT-PCR in various tissues(lung, colon, skeletal muscle, thyroid gland,heart, cecum, spinal cord, and kidney) each isoform of DYNC1LI2 was detected in every tissue. Expression pattern of DYNC1LI2 was validated, and each of DYNC1LI2 transcripts is decreasedafter exercise. By conducting amino acid analysis, our research team investigated cassette exonregion, where phosphorylation site and glycosylation site are exist, and it means, DYNC1LI2acould be more stable in protein between two transcripts. Through this study, DYNC1LI2 genewhich is estimated that is related with exercise has investigated in molecular biological level,and detected data can give warrant as a base data in exercise related researches.
참고문헌 (25)
- Abràmoff MD, "Image processing with Image", J. Biophotonics International, 11(7), pp.36~42 (2004)
- Afzelius BA., "A human syndrome caused by immotile cilia", Science, 193, pp.317~319 (1976)
- Döhner K, "Viral stop-and-go along microtubules: taking a ride with dynein and kinesins", Trends in Microbiology, 13(7), pp.320~327 (2005)
- Doxsey SJ, "Pericentrin, a highly conserved centrosome protein involved in microtubule organization", Cell, 76(4), pp.639~650 (1994)
- Dujardin DL, "A role for cytoplasmic dynein and LIS1 in directed cell movement", The Journal of Cell Biology, 163(6), pp.1205~1211 (2003)
- Fath KR, "Molecular motors are differentially distributed on Golgi membranes from polarized epithelial cells", The Journal of Cell Biology, 126(3), pp.661~675 (1994)
- Fuchs SY, "Phosphorylation-dependent targeting of c-Jun ubiquitination by Jun N-kinase", Oncogene, 13, pp.1531~1535 (1996)
- Hudson RC, "Bioinformatics Analysis of the FREM1 Gene-Evolutionary Development of the IL-1R1 Co-Receptor, TILRR", Biology, 1(3), pp.484~494 (2012)
- Hirokawa N., "Kinesin and dynein superfamily proteins and the mechanism of organelle transport", Science, 279(5350), pp.519~526 (1998)
- Höök P, "The dynein family at a glance", The Journal of Cell Science, 119(21), pp.4369~4371 (2006)
- Kearse M, "Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data", Bioinformatics, 28(12), pp.1647~1649 (2012)
- King SJ, "Analysis of the dynein-dynactin interaction in vitro and in vivo", Molecular Biology of the Cell, 14(12), pp.5089~5097 (2003)
- Lurie M, "Ciliary ultrastructure of respiratory and fallopian tube epithelium in a sterile woman with Kartagener's syndrome. A quantitative estimation", CHEST Journal, 95(3), pp.578~581 (1989)
- Magnani M, "The ubiquitin-dependent proteolytic system and other potential targets for the modulation of nuclear factor-kB(NF-kB)", Curr. Drug Targets, 1, pp.387~399 (2000)
- Mikami A, "Molecular structure of cytoplasmic dynein 2 and its distribution in neuronal and ciliated cells", The Journal of Cell Science, 115(24), pp.4801~4808 (2002)
- Park KD, "Whole transcriptome analyses of six thoroughbred horses before and after exercise using RNA-Seq", BMC Genomics, 13(1), pp.473~ (2012)
- Pazour GJ, "The DHC1b(DHC2) isoform of cytoplasmic dynein is required for flagellar assembly", The Journal of Cell Biology, 144(3), pp.473~481 (1999)
- Porter ME, "Cytoplasmic dynein heavy chain 1b is required for flagellar assembly in Chlamydomonas", Molecular Biology of the Cell, 10(3), pp.693~712 (1999)
- Puls I, "Distal spinal and bulbar muscular atrophy caused by dynactin mutation", Annals of Neurology, 57(5), pp.687~694 (2005)
- Rudd PM, "Roles for glycosylation of cell surface receptors involved in cellular immune recognition", Journal of Molecular Biology, 293(2), pp.351~366 (1999)
- Supp DM, "Mutation of an axonemal dynein affects left– right asymmetry in inversus viscerum mice", Nature, 389(6654), pp.963~966 (1997)
- Tamura K, "MEGA6: Molecular Evolutionary Genetics Analysis version 6.0", Mol. Biol. Evol, 30, pp.2725~2729 (2013)
- Thompson RF, "Myosin superfamily evolutionary history", The Anatomical Record, 268(3), pp.276~289 (2002)
- Treier M, "Ubiquitin-dependent c-Jun degradation in vivo is mediated by the delta domain", Cell, 78, pp.787~798 (1994)
- Vevaina JR, "Correlation of absent inner dynein arms and mucociliary clearance in a patient with Kartagener's syndrome", CHEST Journal, 91(1), pp.91~95 (1987)