title: Regulation of phenylalanine ammonia-lyase (PAL) gene family in wood forming tissue of Populus trichocarpa.
abstract: Phenylalanine ammonia-lyase (PAL) catalyzes the initial step of phenylpropanoid biosynthesis in plants. Five PAL genes (PtrPAL1 to 5) have been identified in Populus trichocarpa. These genes are classified into two subgroups according to their transcript sequence similarity and tissue specificity. However, the regulation of these genes and their protein functions are not well understood. In this study, enzymatic properties of each PtrPALs were characterized based on their recombinant proteins expressed in E.coli. Subcellular localizations of each PtrPALs in stem wood forming tissue were investigated and individual PtrPAL protein abundances in cytosol and membrane protein fractions were measured using protein cleavage-isotope dilution mass spectrometry (PC-IDMS). Protein/mRNA ratios of PtrPALs were further verified using RNA-Seq and gel-enhanced liquid chromatography mass spectrometry (GeLC-MS). All PtrPALs have similar catalytic properties for the deamination of L-phenylalanine, their major substrate. All PtrPALs have similar subcellular locations in stem wood forming tissue, with major amount in the cytosol (93-96 %) and less in the membrane (4-7 %). However, the protein/mRNA ratios of subgroup A (PtrPAL2, 4 and 5) are about five times that of subgroup B (PtrPAL1 and 3) in stem wood forming tissue, while all PtrPALs have similar transcript abundances. These results indicate a greater functional significance of subgroup A PtrPALs for stem wood formation, and highlight the role of gene post-transcriptional regulation.
authors.affiliation: Forest Biotechnology Group, Department of Forestry and Environmental Resources, North Carolina State University, Raleigh, NC 27695, USA. rshi@ncsu.edu
authors.forename: Rui
authors.initials: R
authors.lastname: Shi
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Shuford
authors.forename: Jack P
authors.initials: JP
authors.lastname: Wang
authors.forename: Ying-Hsuan
authors.initials: YH
authors.lastname: Sun
authors.forename: Zhichang
authors.initials: Z
authors.lastname: Yang
authors.forename: Hsi-Chuan
authors.initials: HC
authors.lastname: Chen
authors.forename: Sermsawat
authors.initials: S
authors.lastname: Tunlaya-Anukit
authors.forename: Quanzi
authors.initials: Q
authors.lastname: Li
authors.forename: Jie
authors.initials: J
authors.lastname: Liu
authors.forename: David C
authors.initials: DC
authors.lastname: Muddiman
authors.forename: Ronald R
authors.initials: RR
authors.lastname: Sederoff
authors.forename: Vincent L
authors.initials: VL
authors.lastname: Chiang
mesh_terms: D018506:Gene Expression Regulation, Plant / Q000502:physiology; D013058:Mass Spectrometry; D010650:Phenylalanine Ammonia-Lyase / Q000378:metabolism*; D032107:Populus / Q000201:enzymology* / Q000378:metabolism*
chemical_list: D010650:Phenylalanine Ammonia-Lyase