Dynamin binding protein (Tuba) deficiency inhibits ciliogenesis and nephrogenesis in Vitro and in Vivo

Jeong In Baek, Sang Ho Kwon, Xiaofeng Zuo, Soo Young Choi, Seok Hyung Kim, Joshua H. Lipschutz

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Dysfunction of renal primary cilia leads to polycystic kidney disease. We previously showed that the exocyst, a protein trafficking complex, is essential for ciliogenesis and regulated by multiple Rho and Rab family GTPases, such as Cdc42. Cdc42 deficiency resulted in a disruption of renal ciliogenesis and a polycystic kidney disease phenotype in zebrafish and mice. Here we investigate the role of Dynamin binding protein (also known as Tuba), a Cdc42-specific guanine nucleotide exchange factor, in ciliogenesis and nephrogenesis using Tuba knockdown Madin-Darby canine kidney cells and tuba knockdown in zebrafish. Tuba depletion resulted in an absence of cilia, with impaired apical polarization and inhibition of hepatocyte growth factor-induced tubulogenesis in Tuba knockdown Madin-Darby canine kidney cell cysts cultured in a collagen gel. In zebrafish, tuba was expressed in multiple ciliated organs, and, accordingly, tuba start and splice site morphants showed various ciliary mutant phenotypes in these organs. Co-injection of tuba and cdc42 morpholinos at low doses, which alone had no effect, resulted in genetic synergy and led to abnormal kidney development with highly disorganized pronephric duct cilia. Morpholinos targeting two other guanine nucleotide exchange factors not known to be in the Cdc42/ciliogenesis pathway and a scrambled control morpholino showed no phenotypic effect. Given the molecular nature of Cdc42 and Tuba, our data strongly suggest that tuba and cdc42 act in the same ciliogenesis pathway. Our study demonstrates that Tuba deficiency causes an abnormal renal ciliary and morphogenetic phenotype. Tuba most likely plays a critical role in ciliogenesis and nephrogenesis by regulating Cdc42 activity.

Original languageEnglish (US)
Pages (from-to)8632-8643
Number of pages12
JournalJournal of Biological Chemistry
Volume291
Issue number16
DOIs
StatePublished - Apr 15 2016
Externally publishedYes

Fingerprint

Dynamins
Protein Deficiency
Morpholinos
Guanine Nucleotide Exchange Factors
Cilia
Carrier Proteins
Zebrafish
Kidney
Polycystic Kidney Diseases
Madin Darby Canine Kidney Cells
Phenotype
Hepatocyte Growth Factor
GTP Phosphohydrolases
rab GTP-Binding Proteins
Ducts
Protein Transport
Collagen
Gels
Polarization
Cysts

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Dynamin binding protein (Tuba) deficiency inhibits ciliogenesis and nephrogenesis in Vitro and in Vivo. / Baek, Jeong In; Kwon, Sang Ho; Zuo, Xiaofeng; Choi, Soo Young; Kim, Seok Hyung; Lipschutz, Joshua H.

In: Journal of Biological Chemistry, Vol. 291, No. 16, 15.04.2016, p. 8632-8643.

Research output: Contribution to journalArticle

Baek, Jeong In ; Kwon, Sang Ho ; Zuo, Xiaofeng ; Choi, Soo Young ; Kim, Seok Hyung ; Lipschutz, Joshua H. / Dynamin binding protein (Tuba) deficiency inhibits ciliogenesis and nephrogenesis in Vitro and in Vivo. In: Journal of Biological Chemistry. 2016 ; Vol. 291, No. 16. pp. 8632-8643.
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AU - Lipschutz, Joshua H.

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