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文獻解讀

集思慧遠客戶發表《HY5與組蛋白去乙酰化酶HDA 15相互作用抑制下胚軸細胞在光形態發生中的伸長》

幸运飞艇官方开户FABURIQI:2019-06-24 LIULANCISHU:383

摘要:

GUANGXINGTAIFASHENGSHIYIGEZHONGYAODEZHIWUFAYUGUOCHENG,SHEJIDAOGUANGJIEDAODEZHUANLUTIHEZUDANBAIXIUSHIDEGAIBIAN。XIAPEIZHOUSHENCHANGZHUANLUYINZI5(Hy5)ZUOYONGYUGUANGSHOUTIDUOGEJIAZUDEXIAYOU,TONGGUODIAOJIEGUANGFANYINGDANBAIDEBIAODALAICUJINGUANGXINGTAIDEXINGCHENG。RANER,Hy5JIEDAODEZHUANLUDIAOKONGDEFENZIJIZHISHANGBUQINGCHU。ZAIZHELI,WOMENZHENGMINGHy5ZHIJIEYURPD3/HDA1XINGZUDANBAIQUYIXIANHUAMEIHDA15ZAITIWAIHETINEIXIANGHUZUOYONG。BIAOXINGFENXIBIAOMING,HDA15ZAIHONGGUANGHEYUANHONGGUANGTIAOJIANXIADUININANJIEYOUMIAOXIAPEIZHOUXIBAODESHENCHANGJUYOUFUDIAOJIEZUOYONG。HDA15DEMEIHUOXINGDUIXIAPEIZHOUSHENCHANGYOUYIZHIZUOYONG。CIWAI,HDA 15HEHY5XIANGHUYILAIDIYIZHIXIAPEIZHOUXIBAOZAIGUANGXINGTAIFASHENGZHONGDESHENCHANG。QUANJIYINZUZHUANLUZUFENXIXIANSHIHDA 15HE HY5GONGTONGYIZHIXIBAOBIZUZHIHESHENGCHANGSUXINHAOXIANGGUANJIYINDEZHUANLU。CIWAI,HDA 15ZAIHY5YIZHININANJIEYOUMIAOXIAPEIZHOUXIBAOSHENCHANGXIANGGUANJIYINZHONGDEZUOYONGSHIBIXUDE。CIWAI,HY5HUANJIANGHDA 15ZHAOMUDAOMUBIAOJIYINDEQIDONGZIZHONG,BINGTONGGUOYIGUANGYILAIDEFANGSHIJIANGDIZUDANBAIH4YIXIANHUASHUIPINGLAIYIZHIJIYINDEBIAODA。WOMENDEYANJIUJIESHILEYIGEGUANJIANDEZHUANLUDIAOKONGJIEDIAN,QIZHONGHY5 49YUHDA 15XIANGHUZUOYONG,CANYUYIZHIXIAPEIZHOUXIBAODESHENCHANG,CUJINGUANGXINGTAIFASHENG。

結果

1.HDA 15與HY5相互作用

通過體外和半體外下拉試驗(圖1A及1B)、雙分子熒光互補(BiFC)和共免疫共沉淀(Co-IP)(圖1C及1D),發現HDA15與HY5和HYH直接相互作用。


2.HDA 15是光形態發生中下胚軸伸長的負調節因子

觀察到hda 15突變體hda 15-1在不同光照條件下表現出較長的下胚軸表型。相比之下,35S:HDA15--GFP轉基因株系與野生型(Col-0)在紅光和遠紅光條件下的比較表現出明顯較短的下胚軸表型(圖2A-D)。說明HDA15在下胚軸的光形態發生中具有負作用。為了檢測HDA15與光感受器PHYA和PHYB的遺傳關系,將35s:HDA15--GFP通過雜交導入 Phya-211和phyb-9突變體。

ZHIWUSHENGCHANGSHEJIXIBAODEYANSHENHEZENGZHI,ZUOZHEJINYIBUCEDINGLEHONGGUANGTIAOJIANXIA4RILINGCol-0 HEhad15-1DEXIBAOCHANGDUHEXIBAOSHU。hdda 15-1XIAPEIZHOUDEPINGJUNXIBAOCHANGDUMINGXIANCHANGYUYESHENGXING。XIANGBIZHIXIA,hda 15-1XIBAOSHUYUYESHENGXINGXIBAOSHUWUXIANZHUCHAYI(TU2E-G)ZHEXIESHUJUXIANSHIHDA15ZHUYAODIAOKONGNINANJIEYOUMIAOXIAPEIZHOUXIBAODESHENCHANG。

幸运飞艇官方开户 3.HDA15DEMEIHUOXINGDUIXIAPEIZHOUSHENCHANGDEYIZHIZUOYONG

XIANQIANDEYANJIUBIAOMING,RPD3/HDA1-type HDACs, HDA5, HDA6, HDA15HE HDA19JUYOUZUDANBAIQUYIXIANHUAMEIHUOXING。WOMENJINYIBUCEDINGLEHDA15MEIHUOXINGSUOXUDEANJISUANCANJI。RENHDAC1(HsHDAC 1)HEHDAC8(HsHDAC 8)HENINANJIERPD 3/HDA 1XINGHDACsZUDANBAIJINXINGQUYIXIANHUAMEI(HD)JIEGOUYUDEXULIEBIDUI,BIAOMINGHDACsZHONGDEANJISUANCANJI,H 276,H 277,D 313,H 315HEH 316SHIBAOSHOUDE。WEILEFENXIZHEXIEBAOSHOUDEANJISUANSHIFOUDUIYUHDA15DECUIHUAHUOXINGLAISHUOSHIZHONGYAODE,GOUJIANLEHANYOUANJISUAN(H276A、H277A、D313A、H315AHEH316A)TUBIANDEZHILI,BINGZAIDACHANGGANJUN(E.coli)ZHONGBIAODA。ZUOWEIDUIZHAO,ZAIBUBAOSHOUDEANJISUAN(H282A)SHANGDETUBIANYEPINGXINGFENXI(TU3A)。

4.HD15HEHY5ZAIGUANGXINGTAIFASHENGZHONGYIZHIXIAPEIZHOUSHENCHANGDEXIANGHUZUOYONG

幸运飞艇官方开户 ZUOZHETONGGUOGOUJIANTUBIANTIHEZAJIAO,BINGDUITUBIANTIHEYESHENGXINGZAIHONGGUANGHEYUANHONGGUANGDEBIAOXINGJINXINGCELIANG(TU4)。ZHEXIEJIEGUOBIAOMING,HDA 15HEHY5ZAIGUANGXINGTAIFASHENGGUOCHENGZHONGKENENGXIANGHUYILAIDIDIAOKONGXIAPEIZHOUSHENCHANG。

5.HONGGUANGTIAOJIANXIAHDA15HEHY5GONGDIAOJIYINZHUANLUZUFENXI

幸运飞艇官方开户 YONGmRNA-seqFENXILIhda15HEhy5DIAOJIEZHUANLUZUDEBIANHUA。CAIJI2RILINGZAILIANXUHONGGUANGTIAOJIANXIACOL-0、hda 15-1HEhy5-215YOUMIAO,JINXINGFENXI。YUYESHENGXINGXIANGBI,hda 15HE hy5TUBIANTIFENBIEYOU425HE1134GEJIYINSHANGDIAO,579HE644GEJIYINXIADIAO。CIWAI,hda 15HEHY5GONGDIAOKONGLE499GEJIYIN(TU5A、5B)。HDA15HEHY5GONGDIAOKONGJIYINDEGOFENXI,JIESHIHDA15HEHY5GONGYIZHIJIYINSHIYOUXIANYUDUIFEISHENGWUCIJIDEFANYINGXIANGGUANDE,LIRUDUINEIYUANCIJI、SHUIJIEMEI、SHENGZHIFAYU、XIBAOBIZUZHIHEDUISHENGCHANGSUCIJIDEFANYINGDEFANYING(TU5C)。

幸运飞艇官方开户 6.HDA15HEHY5GONGTONGYIZHIHONGGUANGXIAXIBAOBIHESHENGCHANGSUXINHAOXIANGGUANJIYINBIAODA

RNA-seqSHUJUXIANSHI,HDA15HEHY5GONGYIZHIXIBAOBIZUZHIZHONGSHEJIDEYIXIEJIYIN。RT-qPCRFENXIZHENGSHI,ZAIhda 15HEhy5TUBIANZHONG,XTH10、XTH15、XTH17、EXP 2、BXL1、PME 16HEPME 35DEBIAODASHANGDIAOLE(TU6A),ZHICHI HDA 15HEHY5ZAIXIAPEIZHOUSHENCHANGZHONGDEFUZUOYONG。ZHEXIEJIYINDEZHUANLUWUZAI35S:HDA15-GFPZHONGXIADIAO(TU6B),QUERENHDA15DUICANYUXIAPEIZHOUXIBAOSHENCHANGDEJIYINDEYIZHIZUOYONG。JINYIBUJIANCELEXIBAOBIZUZHIHESHENGCHANGSUXINHAOXIANGGUANJIYINZAIZAIHONGGUANGTIAOJIANXIASHENGCHANG35s:HY5-gfpHE35S:HY5-GFP hda 15YOUMIAOZHONGDEBIAODASHUIPING(TU6C)。

7.HY5ZHAOMUHDA15JIARUXIBAOBIZUZHIHESHENGCHANGSUXINHAOXIANGGUANJIYINDEQIDONGZIZHONG,CHENGGUANGYILAIXING

幸运飞艇官方开户 WEILEQUEDINGZHEXIEXIBAOBIZUZHIHESHENGCHANGSUXINHAOXIANGGUANJIYINSHITINEIHy5DEZHIJIEBADIAN,LIYONGProHY5:HY5-GFPZHUANJIYINZHIZHUBIAODAHY5-GFPYOUTIANRANHY5QIDONGZIQUDONGRANSEZHIMIANYICHENDIAN(CHIP)JIANCE。

幸运飞艇官方开户 8.HY5HEHAD 15JIANGDIBAJIYINZUDANBAIH4YIXIANHUASHUIPING

幸运飞艇官方开户 SinhadHDA15 hJUYOUZUDANBAITUOQINGMEIHUOXING,BINGQIESHIDUIXIAPEIZHOUSHENCHANGDEYIZHISUOBIXUDE。WOMENJINYIBUYANJIUZUDANBAIHDA 15HEHY5GONGDIAOKONGJIYIN、XTH15,EXP 2,IAA 19,COL-0DEH3HEH4SUOQUANHUASHUIPING。

總結

ZAIBENGONGZUOZHONG,QUEDINGHY5YUHDA15ZAITINEIHETIWAIDOUYOUZHIJIEDEXIANGHUZUOYONG。HY5ZHAOMUHDA15,TONGGUOJIANGDININANJIEYOUMIAOZUDANBAIH4YIXIANHUASHUIPING,YIZHIXIBAOBIZUZHIHESHENGCHANGSUXINHAOXIANGGUANDE133GEJIYIN。ZHEXIEFAXIANJIESHILEYIGEGUANJIANDEZHUANLUDIAOKONGJIEDIAN,QIZHONGHY5YUHDA 15XIANGHUZUOYONG,CANYUYIZHIXIAPEIZHOUXIBAOZAIGUANGXINGTAIFASHENGZHONGDESHENCHANG。