Neurabin is a synaptic protein linking p70 S6 kinase and the neuronal cytoskeleton.
p70 S6 kinase (p70(S6k)) is a mitogen-activated protein kinase that plays a central role in the control of mRNA translation. It physiologically phosphorylates the S6 protein of the 40s ribosomal subunit in response to mitogenic stimuli and is a downstream component of the rapamycin-sensitive pathway, which includes the 12-kDa FK506 binding protein and includes rapamycin and the 12-kDa FK506 binding protein target 1. Here, we report the identification of neurabin (neural tissue-specific F-actin binding protein), a neuronally enriched protein of 1,095 amino acids that contains a PDZ domain and binds p70(S6k). We demonstrate the neurabin-p70(S6k) interaction by yeast two-hybrid analysis and biochemical techniques. p70(S6k) and neurabin coimmunoprecipitate from transfected HEK293 cells. Site-directed mutagenesis of neurabin implicates its PDZ domain in the interaction with p70(S6k), and deletion of the carboxyl-terminal five amino acids of p70(S6k) abrogates the interaction. Cotransfection of neurabin in HEK293 cells activates p70(S6k) kinase activity. The mRNA of neurabin and p70(S6k) show striking colocalization in brain sections by in situ hybridization. Subcellular fractionation of rat brain demonstrates that neurabin and p70(S6k) both localize to the soluble fraction of synaptosomes. By way of its PDZ domain, the neuronal-specific neurabin may target p70(S6k) to nerve terminals.
Burnett
P E
PE
Department of Neuroscience, The Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.
Blackshaw
S
S
Lai
M M
MM
Qureshi
I A
IA
Burnett
A F
AF
Sabatini
D M
DM
Snyder
S H
SH
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D003599:Cytoskeleton / Q000378:metabolism; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics*; D006801:Humans; D017403:In Situ Hybridization; D008840:Microfilament Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009419:Nerve Tissue Proteins / Q000235:genetics* / Q000378:metabolism; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure; D051381:Rats; D019893:Ribosomal Protein S6 Kinases / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D013569:Synapses / Q000378:metabolism*; D014162:Transfection; D000099273:Neurabins
D018076:DNA, Complementary; D008840:Microfilament Proteins; D009419:Nerve Tissue Proteins; D019893:Ribosomal Protein S6 Kinases; D000099273:Neurabins