Disruption of phenylalanine hydroxylase reduces adult lifespan and fecundity, and impairs embryonic development in parthenogenetic pea aphids.
Phenylalanine hydroxylase (PAH) is a key tyrosine-biosynthetic enzyme involved in neurological and melanin-associated physiological processes. Despite extensive investigations in holometabolous insects, a PAH contribution to insect embryonic development has never been demonstrated. Here, we have characterized, for the first time, the PAH gene in a hemimetabolous insect, the aphid Acyrthosiphon pisum. Phylogenetic and sequence analyses confirmed that ApPAH is closely related to metazoan PAH, exhibiting the typical ACT regulatory and catalytic domains. Temporal expression patterns suggest that ApPAH has an important role in aphid developmental physiology, its mRNA levels peaking at the end of embryonic development. We used parental dsApPAH treatment to generate successful knockdown in aphid embryos and to study its developmental role. ApPAH inactivation shortens the adult aphid lifespan and considerably affects fecundity by diminishing the number of nymphs laid and impairing embryonic development, with newborn nymphs exhibiting severe morphological defects. Using single nymph HPLC analyses, we demonstrated a significant tyrosine deficiency and a consistent accumulation of the upstream tyrosine precursor, phenylalanine, in defective nymphs, thus confirming the RNAi-mediated disruption of PAH activity. This study provides first insights into the role of PAH in hemimetabolous insects and demonstrates that this metabolic gene is essential for insect embryonic development.
Univ Lyon, INSA-Lyon, INRA, BF2I, UMR0203, F-69621, Villeurbanne, France.
Pierre
P
Simonet
Univ Lyon, INSA-Lyon, INRA, BF2I, UMR0203, F-69621, Villeurbanne, France.
Karen
K
Gaget
Univ Lyon, INSA-Lyon, INRA, BF2I, UMR0203, F-69621, Villeurbanne, France.
Nicolas
N
Parisot
Univ Lyon, INSA-Lyon, INRA, BF2I, UMR0203, F-69621, Villeurbanne, France.
Gabrielle
G
Duport
Univ Lyon, INSA-Lyon, INRA, BF2I, UMR0203, F-69621, Villeurbanne, France.
Marjolaine
M
Rey
Univ Lyon, INSA-Lyon, INRA, BF2I, UMR0203, F-69621, Villeurbanne, France.
Gérard
G
Febvay
Univ Lyon, INSA-Lyon, INRA, BF2I, UMR0203, F-69621, Villeurbanne, France.
Hubert
H
Charles
KU Leuven, University of Leuven, Department of Human Genetics, Laboratory of Behavioral and Developmental Genetics, B-3000, Leuven, Belgium.|VIB Center for the Biology of Disease, B-3000, Leuven, Belgium.
Patrick
P
Callaerts
Univ Lyon, INSA-Lyon, INRA, BF2I, UMR0203, F-69621, Villeurbanne, France.
Stefano
S
Colella
Univ Lyon, INSA-Lyon, INRA, BF2I, UMR0203, F-69621, Villeurbanne, France.
Federica
F
Calevro
D000818:Animals; D001042:Aphids / Q000196:embryology* / Q000502:physiology; D005298:Fertility; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D008136:Longevity; D010312:Parthenogenesis; D018532:Pisum sativum / Q000469:parasitology*; D010651:Phenylalanine Hydroxylase / Q000235:genetics*; D010802:Phylogeny
D010651:Phenylalanine Hydroxylase
Aging Dis. 2015 Oct 01;6(5):390-9
26425393
PLoS One. 2013 Nov 14;8(11):e79482
24244510
Biochem J. 1990 Apr 15;267(2):399-405
2334400
Adv Genet. 1987;24:127-222
3124532
Nature. 1999 Sep 30;401(6752):447-52
10519548
Biochemistry. 1985 Jan 29;24(3):556-61
2986678
PLoS One. 2013 Aug 26;8(8):e71984
23991017
Insect Mol Biol. 2010 Mar;19 Suppl 2:5-12
20482635
J Biol Chem. 1986 Mar 25;261(9):4148-53
2869038
Trends Parasitol. 2005 Apr;21(4):192-9
15780842
Science. 2014 Nov 7;346(6210):763-7
25378627
Nucleic Acids Res. 2015 Jan;43(Database issue):D222-6
25414356
Cell Mol Life Sci. 2011 Apr;68(8):1297-309
21390549
Proc Natl Acad Sci U S A. 2011 Feb 15;108(7):2849-54
21282658
J Neurogenet. 1999 Nov;13(3):157-80
10928216
Dev Comp Immunol. 1998 Sep-Dec;22(5-6):469-78
9877430
Mol Biol Evol. 2010 Feb;27(2):221-4
19854763
PLoS One. 2014 Jan 07;9(1):e84865
24409310
PLoS One. 2012;7(9):e46209
23049981
Mol Ecol. 2014 Mar;23(6):1473-96
23952067
IUBMB Life. 2013 Apr;65(4):341-9
23457044
Insect Biochem Mol Biol. 2014 Aug;51:20-32
24859468
Nucleic Acids Res. 2004 Mar 19;32(5):1792-7
15034147
FASEB J. 2008 Aug;22(8):3046-58
18460651
Clin Biochem Rev. 2008 Feb;29(1):31-41
18566668
Insect Biochem Mol Biol. 2004 Dec;34(12):1329-38
15544946
J Biol Chem. 2007 Jul 27;282(30):21973-86
17537732
Biotechnol Lett. 2004 Mar;26(6):509-15
15127793
Biochem J. 1995 Oct 15;311 ( Pt 2):353-66
7487868
J Cereb Blood Flow Metab. 2008 Apr;28(4):824-31
17971791
Gene. 2008 Dec 31;427(1-2):86-92
18835579
Mol Genet Metab. 2014 Apr;111(4):452-61
24560888
Insect Mol Biol. 2010 Mar;19 Suppl 2:249-58
20482655
Insect Biochem Mol Biol. 1994 Jun;24(6):581-8
7913853
J Mol Biol. 2002 Jul 12;320(3):645-61
12096915
Proc Natl Acad Sci U S A. 2000 Oct 10;97(21):11397-402
11005829
Mol Biochem Parasitol. 2011 Jan;175(1):58-67
20887755
PLoS Biol. 2010 Feb 23;8(2):e1000313
20186266
Insect Biochem Mol Biol. 2003 Mar;33(3):345-54
12609519
JIMD Rep. 2015;20:35-8
25614310
Microbiol Mol Biol Rev. 2004 Dec;68(4):745-70
15590782
Curr Opin Genet Dev. 2010 Aug;20(4):355-61
20462751
Appl Environ Microbiol. 2013 May;79(10):3225-33
23503309
Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1366-70
8108417
Sci Rep. 2016 Jan 29;6:19967
26822159
Hum Mutat. 2007 Sep;28(9):831-45
17443661
Nat Struct Biol. 1999 May;6(5):401-2
10331859
BMC Genomics. 2013 Apr 10;14:235
23575215
Insect Sci. 2014 Feb;21(1):13-9
23955993
Gene. 1990 Sep 14;93(2):213-9
2121612
Biochem Genet. 1980 Aug;18(7-8):781-91
6781472
Plant Cell. 2010 Oct;22(10):3410-22
20959559
J Exp Zool B Mol Dev Evol. 2003 Feb 15;295(1):59-81
12548543
Nucleic Acids Res. 2010 Jul;38(Web Server issue):W332-9
20444868
Syst Biol. 2010 May;59(3):307-21
20525638
Biochem J. 1992 Oct 1;287 ( Pt 1):85-9
1417795