Small interfering peptide (siP) for in vivo examination of the developing lung interactonome.
To understand the role of reactive oxygen species in mechanosensory control of lung development a new approach to interfere with protein-protein interactions by means of a short interacting peptide was developed. This technology was used in the developing rodent lung to examine the role of NADPH oxidase (NOX), casein kinase 2 (CK2), and the cystic fibrosis transmembrane conductance regulator (CFTR) in stretch-induced differentiation. Interactions between these molecules was targeted in an in utero system with recombinant adeno-associated virus (rAAV) containing inserted DNA sequences that express a control peptide or small interfering peptides (siPs) specific for subunit interaction or phosphorylation predicted to be necessary for multimeric enzyme formation. In all cases only siPs with sequences necessary for a predicted normal function were found to interfere with assembly of the multimeric enzyme. A noninterfering control siP to nonessential regions or reporter genes alone had no effect. Physiologically, it was shown that siPs that interfered with the NOX-CFTR-CK2 complex that we call an "interactonome" affected markers of stretch-induced lung organogenesis including Wnt/beta-catenin signaling.
Cohen
J Craig
JC
The Brady Laboratory, Section of Neonatology, Department of Pediatrics, Stony Brook University, School of Medicine, Stony Brook, New York 11794, USA. jcraig.cohen@stonybrook.edu
Killeen
Erin
E
Chander
Avinash
A
Takemaru
Ken-Ichi
K
Larson
Janet E
JE
Treharne
Kate J
KJ
Mehta
Anil
A
D000595:Amino Acid Sequence; D000818:Animals; D047390:Casein Kinase II / Q000235:genetics / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics; D008168:Lung / Q000196:embryology / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D055503:Protein Multimerization; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction; D051153:Wnt Proteins / Q000502:physiology; D051176:beta Catenin / Q000502:physiology
D010455:Peptides; D017382:Reactive Oxygen Species; D051153:Wnt Proteins; D051176:beta Catenin; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D019255:NADPH Oxidases; D047390:Casein Kinase II