Generation and Characterization of a Specific Polyclonal Antibody against Arabidopsis thaliana Phytochrome-Interacting Factor 3

被引：0
作者：
Da-Min Choi
Jae-Yong Cho
Woe-Yeon Kim
Yun-Jeong Han
Jeong-Il Kim
机构：
[1] Chonnam National University,Department of Integrative Food, Bioscience and Biotechnology
[2] Gyeongsang National University,Department of Applied Life Chemistry
[3] Kumho Life Science Laboratory,undefined
[4] Chonnam National University,undefined
来源：
Journal of Plant Biology | 2021年 / 64卷
关键词：
Phytochrome; Phytochrome-interacting factors; PIF3; Polyclonal antibody; Protein degradation;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
Phytochrome-interacting factors (PIFs) are a basic helix-loop-helix family of transcriptional regulators that maintain skotomorphogenesis and suppress photomorphogenesis. PIFs are regulated by plant photoreceptors, especially phytochromes. In general, PIFs physically interact with phytochromes, and this interaction induces PIF’s phosphorylation and subsequent degradation, contributing to the initiation of photomorphogenic development. Among the eight members of PIF (PIF1 to PIF8) reported in Arabidopsis thaliana, PIF3 is the first discovered member and plays central roles in de-etiolation and chlorophyll biosynthesis. More recently, PIF3 has been also reported to regulate hormone signaling and cold tolerance in plants. Although PIF3 protein shows dynamic behaviors in plants, its study has been limited due to the lack of an authentic PIF3 antibody. In this study, we produced polyclonal antibodies using inclusion bodies and characterized the PIF3 antibody based on specificity and sensitivity. In addition, we investigated PIF3 phosphorylation and degradation during phytochrome-mediated light signaling in plants. Furthermore, we successfully performed in vitro protein–protein interaction and co-immunoprecipitation assays between phytochrome B (phyB) and PIF3 using the antibody. Therefore, we obtained an authentic PIF3 antibody that could be used as a valuable tool to study the multi-faceted functions of PIF3.
引用
收藏
页码：181 / 191
页数：10
相关论文
共 225 条
  • [1] Adam E(2013)Comparative functional analysis of full-length and N-terminal fragments of phytochrome C, D and E in red light-induced signaling New Phytol 200 86-96
  • [2] Kircher S(2006)Photoactivated phytochrome induces rapid PIF3 phosphorylation prior to proteasome-mediated degradation Mol Cell 23 439-446
  • [3] Liu P(2008)Decoding of light signals by plant phytochromes and their interacting proteins Annu Rev Plant Biol 59 281-311
  • [4] Merai Z(2015)Thermoperiodic control of hypocotyl elongation depends on auxin-induced ethylene signaling that controls downstream PHYTOCHROME INTERACTING FACTOR3 activity Plant Physiol 167 517-530
  • [5] Gonzalez-Schain N(2013)Photoreceptor signaling networks in plant responses to shade Annu Rev Plant Biol 64 403-427
  • [6] Horner M(2013)Antagonistic basic helix-loop-helix/bZIP transcription factors form transcriptional modules that integrate light and reactive oxygen species signaling in Arabidopsis Plant Cell 25 1657-1673
  • [7] Viczian A(2019)The transcription factors TCP4 and PIF3 antagonistically regulate organ-specific light induction of Plant Cell 31 1155-1170
  • [8] Monte E(2020) genes to modulate cotyledon opening during de-etiolation in Arabidopsis Physiol Plant 169 325-335
  • [9] Sharrock RA(2005)Mechanisms regulating PIF transcription factor activity at the protein level Ann Bot 96 169-175
  • [10] Schafer E(2003)Phytochromes and shade-avoidance responses in plants Plant Cell 15 1981-1989
12345678910