Mutation of a barrier insulator in the human ankyrin-1 gene is associated with hereditary spherocytosis

Patrick G. Gallagher, Laurie A. Steiner, Robert I. Liem, Ashley N. Owen, Amanda P. Cline, Nancy E. Seidel, Lisa J. Garrett, David M. Bodine

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Defects of the ankyrin-1 gene are the most common cause in humans of hereditary spherocytosis, an inherited anemia that affects patients of all ethnic groups. In some kindreds, linked -108/-153 nucleotide substitutions have been found in the upstream region of the ankyrin gene promoter that is active in erythroid cells. In vivo, the ankyrin erythroid promoter and its upstream region direct position-independent, uniform expression, a property of barrier insulators. Using human erythroid cell lines and primary cells and transgenic mice, here we have demonstrated that a region upstream of the erythroid promoter is a barrier insulator in vivo in erythroid cells. The region exhibited both functional and structural characteristics of a barrier, including prevention of gene silencing in an in vivo functional assay, appropriate chromatin configuration, and occupancy by barrier-associated proteins. Fragments with the -108/-153 spherocytosis-associated mutations failed to function as barrier insulators in vivo and demonstrated perturbations in barrier-associated chromatin configuration. In transgenic mice, flanking a mutant -108/-153 ankyrin gene promoter with the well-characterized chicken HS4 barrier insulator restored position-independent, uniform expression at levels comparable to wild-type. These data indicate that an upstream region of the ankyrin-1 erythroid promoter acts as a barrier insulator and identify disruption of the barrier element as a potential pathogenetic mechanism of human disease.

Original languageEnglish (US)
Pages (from-to)4453-4465
Number of pages13
JournalJournal of Clinical Investigation
Volume120
Issue number12
DOIs
StatePublished - Dec 1 2010

Fingerprint

Hereditary Spherocytosis
Ankyrins
Erythroid Cells
Mutation
Genes
Transgenic Mice
Chromatin
Gene Silencing
Ethnic Groups
Anemia
Chickens
Nucleotides
Cell Line
Proteins

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Gallagher, P. G., Steiner, L. A., Liem, R. I., Owen, A. N., Cline, A. P., Seidel, N. E., ... Bodine, D. M. (2010). Mutation of a barrier insulator in the human ankyrin-1 gene is associated with hereditary spherocytosis. Journal of Clinical Investigation, 120(12), 4453-4465. https://doi.org/10.1172/JCI42240
Gallagher, Patrick G. ; Steiner, Laurie A. ; Liem, Robert I. ; Owen, Ashley N. ; Cline, Amanda P. ; Seidel, Nancy E. ; Garrett, Lisa J. ; Bodine, David M. / Mutation of a barrier insulator in the human ankyrin-1 gene is associated with hereditary spherocytosis. In: Journal of Clinical Investigation. 2010 ; Vol. 120, No. 12. pp. 4453-4465.
@article{65b8f16788b04c3e86f19784673d5833,
title = "Mutation of a barrier insulator in the human ankyrin-1 gene is associated with hereditary spherocytosis",
abstract = "Defects of the ankyrin-1 gene are the most common cause in humans of hereditary spherocytosis, an inherited anemia that affects patients of all ethnic groups. In some kindreds, linked -108/-153 nucleotide substitutions have been found in the upstream region of the ankyrin gene promoter that is active in erythroid cells. In vivo, the ankyrin erythroid promoter and its upstream region direct position-independent, uniform expression, a property of barrier insulators. Using human erythroid cell lines and primary cells and transgenic mice, here we have demonstrated that a region upstream of the erythroid promoter is a barrier insulator in vivo in erythroid cells. The region exhibited both functional and structural characteristics of a barrier, including prevention of gene silencing in an in vivo functional assay, appropriate chromatin configuration, and occupancy by barrier-associated proteins. Fragments with the -108/-153 spherocytosis-associated mutations failed to function as barrier insulators in vivo and demonstrated perturbations in barrier-associated chromatin configuration. In transgenic mice, flanking a mutant -108/-153 ankyrin gene promoter with the well-characterized chicken HS4 barrier insulator restored position-independent, uniform expression at levels comparable to wild-type. These data indicate that an upstream region of the ankyrin-1 erythroid promoter acts as a barrier insulator and identify disruption of the barrier element as a potential pathogenetic mechanism of human disease.",
author = "Gallagher, {Patrick G.} and Steiner, {Laurie A.} and Liem, {Robert I.} and Owen, {Ashley N.} and Cline, {Amanda P.} and Seidel, {Nancy E.} and Garrett, {Lisa J.} and Bodine, {David M.}",
year = "2010",
month = "12",
day = "1",
doi = "10.1172/JCI42240",
language = "English (US)",
volume = "120",
pages = "4453--4465",
journal = "Journal of Clinical Investigation",
issn = "0021-9738",
publisher = "The American Society for Clinical Investigation",
number = "12",

}

Gallagher, PG, Steiner, LA, Liem, RI, Owen, AN, Cline, AP, Seidel, NE, Garrett, LJ & Bodine, DM 2010, 'Mutation of a barrier insulator in the human ankyrin-1 gene is associated with hereditary spherocytosis', Journal of Clinical Investigation, vol. 120, no. 12, pp. 4453-4465. https://doi.org/10.1172/JCI42240

Mutation of a barrier insulator in the human ankyrin-1 gene is associated with hereditary spherocytosis. / Gallagher, Patrick G.; Steiner, Laurie A.; Liem, Robert I.; Owen, Ashley N.; Cline, Amanda P.; Seidel, Nancy E.; Garrett, Lisa J.; Bodine, David M.

In: Journal of Clinical Investigation, Vol. 120, No. 12, 01.12.2010, p. 4453-4465.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Mutation of a barrier insulator in the human ankyrin-1 gene is associated with hereditary spherocytosis

AU - Gallagher, Patrick G.

AU - Steiner, Laurie A.

AU - Liem, Robert I.

AU - Owen, Ashley N.

AU - Cline, Amanda P.

AU - Seidel, Nancy E.

AU - Garrett, Lisa J.

AU - Bodine, David M.

PY - 2010/12/1

Y1 - 2010/12/1

N2 - Defects of the ankyrin-1 gene are the most common cause in humans of hereditary spherocytosis, an inherited anemia that affects patients of all ethnic groups. In some kindreds, linked -108/-153 nucleotide substitutions have been found in the upstream region of the ankyrin gene promoter that is active in erythroid cells. In vivo, the ankyrin erythroid promoter and its upstream region direct position-independent, uniform expression, a property of barrier insulators. Using human erythroid cell lines and primary cells and transgenic mice, here we have demonstrated that a region upstream of the erythroid promoter is a barrier insulator in vivo in erythroid cells. The region exhibited both functional and structural characteristics of a barrier, including prevention of gene silencing in an in vivo functional assay, appropriate chromatin configuration, and occupancy by barrier-associated proteins. Fragments with the -108/-153 spherocytosis-associated mutations failed to function as barrier insulators in vivo and demonstrated perturbations in barrier-associated chromatin configuration. In transgenic mice, flanking a mutant -108/-153 ankyrin gene promoter with the well-characterized chicken HS4 barrier insulator restored position-independent, uniform expression at levels comparable to wild-type. These data indicate that an upstream region of the ankyrin-1 erythroid promoter acts as a barrier insulator and identify disruption of the barrier element as a potential pathogenetic mechanism of human disease.

AB - Defects of the ankyrin-1 gene are the most common cause in humans of hereditary spherocytosis, an inherited anemia that affects patients of all ethnic groups. In some kindreds, linked -108/-153 nucleotide substitutions have been found in the upstream region of the ankyrin gene promoter that is active in erythroid cells. In vivo, the ankyrin erythroid promoter and its upstream region direct position-independent, uniform expression, a property of barrier insulators. Using human erythroid cell lines and primary cells and transgenic mice, here we have demonstrated that a region upstream of the erythroid promoter is a barrier insulator in vivo in erythroid cells. The region exhibited both functional and structural characteristics of a barrier, including prevention of gene silencing in an in vivo functional assay, appropriate chromatin configuration, and occupancy by barrier-associated proteins. Fragments with the -108/-153 spherocytosis-associated mutations failed to function as barrier insulators in vivo and demonstrated perturbations in barrier-associated chromatin configuration. In transgenic mice, flanking a mutant -108/-153 ankyrin gene promoter with the well-characterized chicken HS4 barrier insulator restored position-independent, uniform expression at levels comparable to wild-type. These data indicate that an upstream region of the ankyrin-1 erythroid promoter acts as a barrier insulator and identify disruption of the barrier element as a potential pathogenetic mechanism of human disease.

UR - http://www.scopus.com/inward/record.url?scp=78649819161&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78649819161&partnerID=8YFLogxK

U2 - 10.1172/JCI42240

DO - 10.1172/JCI42240

M3 - Article

C2 - 21099109

AN - SCOPUS:78649819161

VL - 120

SP - 4453

EP - 4465

JO - Journal of Clinical Investigation

JF - Journal of Clinical Investigation

SN - 0021-9738

IS - 12

ER -