Aberrant splicing in the PKD2 gene as a cause of polycystic kidney disease

David M. Reynolds, Tomohito Hayashi, Yiqiang Cai, Barbera Veldhuisen, Terry J. Watnick, Xose M. Lens, Toshio Mochizuki, Feng Qian, Yoshiko Maeda, Li Li, Ragnheidur Fossdal, Eliecer Coto, Guanqing Wu, Martijn H. Breuning, Gregory G. Germino, Dorien J M Peters, Stefan Somlo

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

It is estimated that approximately 15% of families with autosomal dominant polycystic kidney disease (ADPKD) have mutations in PKD2. Identification of these mutations is central to identifying functionally important regions of gene and to understanding the mechanisms underlying the pathogenesis of the disorder. The current study describes mutations in six type 2 ADPKD families. Two single base substitution mutations discovered in the ORF in exon 14 constitute the most COOH-terminal pathogenic variants described to date. One of these mutations is a nonsense change and the other encodes an apparent missense variant. Reverse transcription-PCR from patient lymphoblast RNA showed that, in addition, both mutations resulted in out-of- frame splice variants by activating cryptic splice sites via different mechanisms. The apparent missense variant produced such a strong splicing signal that the processed transcript from the mutant chromosome did not contain any of the normally spliced, missense product. A third mutation, a nonconservative missense change effecting a negatively charged residue in the third transmembrane span, is likely pathogenic and defines a highly conserved residue consistent with a potential channel subunit function for polycystin- 2. The remaining three mutations included two frame shifts resulting from deletion of one or two bases in exons 6 and 10, respectively, and a nonsense mutation due to a single base substitution in exon 4. The study also defined a novel intragenic polymorphism in exon 1 that will be useful in analyzing 'second hits' in PKD2. Finally, the study demonstrates that there are reduced levels of normal polycystin-2 protein in lymphoblast lines from PKD2-affected individuals and that truncated mutant polycystin-2 cannot be detected in patient lymphoblasts, suggesting that the latter may be unstable in at least some tissues. The mutations described will serve as critical reagents for future functional studies in PKD2.

Original languageEnglish (US)
Pages (from-to)2342-2351
Number of pages10
JournalJournal of the American Society of Nephrology
Volume10
Issue number11
StatePublished - Nov 1999

Fingerprint

Polycystic Kidney Diseases
Mutation
Genes
Exons
Autosomal Dominant Polycystic Kidney
RNA Splice Sites
Nonsense Codon
Open Reading Frames
Reverse Transcription
Chromosomes
RNA
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Nephrology

Cite this

Reynolds, D. M., Hayashi, T., Cai, Y., Veldhuisen, B., Watnick, T. J., Lens, X. M., ... Somlo, S. (1999). Aberrant splicing in the PKD2 gene as a cause of polycystic kidney disease. Journal of the American Society of Nephrology, 10(11), 2342-2351.

Aberrant splicing in the PKD2 gene as a cause of polycystic kidney disease. / Reynolds, David M.; Hayashi, Tomohito; Cai, Yiqiang; Veldhuisen, Barbera; Watnick, Terry J.; Lens, Xose M.; Mochizuki, Toshio; Qian, Feng; Maeda, Yoshiko; Li, Li; Fossdal, Ragnheidur; Coto, Eliecer; Wu, Guanqing; Breuning, Martijn H.; Germino, Gregory G.; Peters, Dorien J M; Somlo, Stefan.

In: Journal of the American Society of Nephrology, Vol. 10, No. 11, 11.1999, p. 2342-2351.

Research output: Contribution to journalArticle

Reynolds, DM, Hayashi, T, Cai, Y, Veldhuisen, B, Watnick, TJ, Lens, XM, Mochizuki, T, Qian, F, Maeda, Y, Li, L, Fossdal, R, Coto, E, Wu, G, Breuning, MH, Germino, GG, Peters, DJM & Somlo, S 1999, 'Aberrant splicing in the PKD2 gene as a cause of polycystic kidney disease', Journal of the American Society of Nephrology, vol. 10, no. 11, pp. 2342-2351.
Reynolds DM, Hayashi T, Cai Y, Veldhuisen B, Watnick TJ, Lens XM et al. Aberrant splicing in the PKD2 gene as a cause of polycystic kidney disease. Journal of the American Society of Nephrology. 1999 Nov;10(11):2342-2351.
Reynolds, David M. ; Hayashi, Tomohito ; Cai, Yiqiang ; Veldhuisen, Barbera ; Watnick, Terry J. ; Lens, Xose M. ; Mochizuki, Toshio ; Qian, Feng ; Maeda, Yoshiko ; Li, Li ; Fossdal, Ragnheidur ; Coto, Eliecer ; Wu, Guanqing ; Breuning, Martijn H. ; Germino, Gregory G. ; Peters, Dorien J M ; Somlo, Stefan. / Aberrant splicing in the PKD2 gene as a cause of polycystic kidney disease. In: Journal of the American Society of Nephrology. 1999 ; Vol. 10, No. 11. pp. 2342-2351.
@article{95fbb2e89b8b4f55aa3c607d65b8563f,
title = "Aberrant splicing in the PKD2 gene as a cause of polycystic kidney disease",
abstract = "It is estimated that approximately 15{\%} of families with autosomal dominant polycystic kidney disease (ADPKD) have mutations in PKD2. Identification of these mutations is central to identifying functionally important regions of gene and to understanding the mechanisms underlying the pathogenesis of the disorder. The current study describes mutations in six type 2 ADPKD families. Two single base substitution mutations discovered in the ORF in exon 14 constitute the most COOH-terminal pathogenic variants described to date. One of these mutations is a nonsense change and the other encodes an apparent missense variant. Reverse transcription-PCR from patient lymphoblast RNA showed that, in addition, both mutations resulted in out-of- frame splice variants by activating cryptic splice sites via different mechanisms. The apparent missense variant produced such a strong splicing signal that the processed transcript from the mutant chromosome did not contain any of the normally spliced, missense product. A third mutation, a nonconservative missense change effecting a negatively charged residue in the third transmembrane span, is likely pathogenic and defines a highly conserved residue consistent with a potential channel subunit function for polycystin- 2. The remaining three mutations included two frame shifts resulting from deletion of one or two bases in exons 6 and 10, respectively, and a nonsense mutation due to a single base substitution in exon 4. The study also defined a novel intragenic polymorphism in exon 1 that will be useful in analyzing 'second hits' in PKD2. Finally, the study demonstrates that there are reduced levels of normal polycystin-2 protein in lymphoblast lines from PKD2-affected individuals and that truncated mutant polycystin-2 cannot be detected in patient lymphoblasts, suggesting that the latter may be unstable in at least some tissues. The mutations described will serve as critical reagents for future functional studies in PKD2.",
author = "Reynolds, {David M.} and Tomohito Hayashi and Yiqiang Cai and Barbera Veldhuisen and Watnick, {Terry J.} and Lens, {Xose M.} and Toshio Mochizuki and Feng Qian and Yoshiko Maeda and Li Li and Ragnheidur Fossdal and Eliecer Coto and Guanqing Wu and Breuning, {Martijn H.} and Germino, {Gregory G.} and Peters, {Dorien J M} and Stefan Somlo",
year = "1999",
month = "11",
language = "English (US)",
volume = "10",
pages = "2342--2351",
journal = "Journal of the American Society of Nephrology : JASN",
issn = "1046-6673",
publisher = "American Society of Nephrology",
number = "11",

}

TY - JOUR

T1 - Aberrant splicing in the PKD2 gene as a cause of polycystic kidney disease

AU - Reynolds, David M.

AU - Hayashi, Tomohito

AU - Cai, Yiqiang

AU - Veldhuisen, Barbera

AU - Watnick, Terry J.

AU - Lens, Xose M.

AU - Mochizuki, Toshio

AU - Qian, Feng

AU - Maeda, Yoshiko

AU - Li, Li

AU - Fossdal, Ragnheidur

AU - Coto, Eliecer

AU - Wu, Guanqing

AU - Breuning, Martijn H.

AU - Germino, Gregory G.

AU - Peters, Dorien J M

AU - Somlo, Stefan

PY - 1999/11

Y1 - 1999/11

N2 - It is estimated that approximately 15% of families with autosomal dominant polycystic kidney disease (ADPKD) have mutations in PKD2. Identification of these mutations is central to identifying functionally important regions of gene and to understanding the mechanisms underlying the pathogenesis of the disorder. The current study describes mutations in six type 2 ADPKD families. Two single base substitution mutations discovered in the ORF in exon 14 constitute the most COOH-terminal pathogenic variants described to date. One of these mutations is a nonsense change and the other encodes an apparent missense variant. Reverse transcription-PCR from patient lymphoblast RNA showed that, in addition, both mutations resulted in out-of- frame splice variants by activating cryptic splice sites via different mechanisms. The apparent missense variant produced such a strong splicing signal that the processed transcript from the mutant chromosome did not contain any of the normally spliced, missense product. A third mutation, a nonconservative missense change effecting a negatively charged residue in the third transmembrane span, is likely pathogenic and defines a highly conserved residue consistent with a potential channel subunit function for polycystin- 2. The remaining three mutations included two frame shifts resulting from deletion of one or two bases in exons 6 and 10, respectively, and a nonsense mutation due to a single base substitution in exon 4. The study also defined a novel intragenic polymorphism in exon 1 that will be useful in analyzing 'second hits' in PKD2. Finally, the study demonstrates that there are reduced levels of normal polycystin-2 protein in lymphoblast lines from PKD2-affected individuals and that truncated mutant polycystin-2 cannot be detected in patient lymphoblasts, suggesting that the latter may be unstable in at least some tissues. The mutations described will serve as critical reagents for future functional studies in PKD2.

AB - It is estimated that approximately 15% of families with autosomal dominant polycystic kidney disease (ADPKD) have mutations in PKD2. Identification of these mutations is central to identifying functionally important regions of gene and to understanding the mechanisms underlying the pathogenesis of the disorder. The current study describes mutations in six type 2 ADPKD families. Two single base substitution mutations discovered in the ORF in exon 14 constitute the most COOH-terminal pathogenic variants described to date. One of these mutations is a nonsense change and the other encodes an apparent missense variant. Reverse transcription-PCR from patient lymphoblast RNA showed that, in addition, both mutations resulted in out-of- frame splice variants by activating cryptic splice sites via different mechanisms. The apparent missense variant produced such a strong splicing signal that the processed transcript from the mutant chromosome did not contain any of the normally spliced, missense product. A third mutation, a nonconservative missense change effecting a negatively charged residue in the third transmembrane span, is likely pathogenic and defines a highly conserved residue consistent with a potential channel subunit function for polycystin- 2. The remaining three mutations included two frame shifts resulting from deletion of one or two bases in exons 6 and 10, respectively, and a nonsense mutation due to a single base substitution in exon 4. The study also defined a novel intragenic polymorphism in exon 1 that will be useful in analyzing 'second hits' in PKD2. Finally, the study demonstrates that there are reduced levels of normal polycystin-2 protein in lymphoblast lines from PKD2-affected individuals and that truncated mutant polycystin-2 cannot be detected in patient lymphoblasts, suggesting that the latter may be unstable in at least some tissues. The mutations described will serve as critical reagents for future functional studies in PKD2.

UR - http://www.scopus.com/inward/record.url?scp=0032736231&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032736231&partnerID=8YFLogxK

M3 - Article

C2 - 10541293

AN - SCOPUS:0032736231

VL - 10

SP - 2342

EP - 2351

JO - Journal of the American Society of Nephrology : JASN

JF - Journal of the American Society of Nephrology : JASN

SN - 1046-6673

IS - 11

ER -