Common traffic routes for imported spermine and endosomal glypican-1-derived heparan sulfate in fibroblasts
Cheng, Fang LU ; Fransson, Lars Åke LU and Mani, Katrin LU
(2018)
In Experimental Cell Research
364(2).
p.133-142
- Abstract
Import of the polyamine spermine from the extracellular environment depends on the presence of cell surface heparan sulfate proteoglycans, such as glypican-1. This proteoglycan is internalized by endocytosis, releases its heparan sulfate chains in endosomes by a nitric oxide-, copper- and amyloid precursor protein-dependent mechanism, then penetrates the membrane and is transported to the nucleus and then to autophagosomes. This process is spontaneous or induced by ascorbate depending on the growth-state of the cell. Here, we have explored possible connections between the heparan sulfate traffic route and spermine uptake and delivery in wild-type and Tg2576 mouse fibroblasts. Cells were examined by deconvolution immunofluorescence... (More)
Import of the polyamine spermine from the extracellular environment depends on the presence of cell surface heparan sulfate proteoglycans, such as glypican-1. This proteoglycan is internalized by endocytosis, releases its heparan sulfate chains in endosomes by a nitric oxide-, copper- and amyloid precursor protein-dependent mechanism, then penetrates the membrane and is transported to the nucleus and then to autophagosomes. This process is spontaneous or induced by ascorbate depending on the growth-state of the cell. Here, we have explored possible connections between the heparan sulfate traffic route and spermine uptake and delivery in wild-type and Tg2576 mouse fibroblasts. Cells were examined by deconvolution immunofluorescence microscopy. The antibodies used were specific for spermine, glypican-1-derived heparan sulfate, Rab7, nucleolin and a marker for autophagosomes. Endogenous immunostainable spermine was primarily associated with autophagosomes. When spermine synthesis was inhibited, imported spermine appeared in Rab7-positive endosomes. When ascorbate was added, heparan sulfate and spermine were transported to the nucleus where they colocalized with nucleolin. Spermine also appeared in autophagosomes. In a pulse-chase experiment, heparan sulfate and spermine were first arrested in late endosomes by actinomycin D treatment. During the chase, when arrest was abolished, heparan sulfate and spermine were both transported to the nucleus and targeted nucleolin. In amyloid precursor protein-/--fibroblasts, ascorbate failed to induce release of heparan sulfate and spermine remained in the endosomes. We propose that cell surface glypican-1 carries spermine to the endosomes and that the released heparan sulfate carries spermine across the membrane into the cytosol and then to the nucleus.
(Less)
- author
- Cheng, Fang
LU
; Fransson, Lars Åke
LU
and Mani, Katrin
LU
- organization
- publishing date
- 2018-02-08
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Amyloid precursor protein, Ascorbate, Autophagosomes, Glypican-1, Heparan sulfate, Nitric oxide, Nucleolin, Spermine
- in
- Experimental Cell Research
- volume
- 364
- issue
- 2
- pages
- 133 - 142
- publisher
- Academic Press
- external identifiers
-
- pmid:29408503
- scopus:85041647261
- ISSN
- 0014-4827
- DOI
- 10.1016/j.yexcr.2018.01.029
- language
- English
- LU publication?
- yes
- id
- f484212f-1b5b-4cfa-a48b-ad0571f083d8
- date added to LUP
- 2018-02-21 14:38:36
- date last changed
- 2024-03-18 05:29:46
@article{f484212f-1b5b-4cfa-a48b-ad0571f083d8,
abstract = {{<p>Import of the polyamine spermine from the extracellular environment depends on the presence of cell surface heparan sulfate proteoglycans, such as glypican-1. This proteoglycan is internalized by endocytosis, releases its heparan sulfate chains in endosomes by a nitric oxide-, copper- and amyloid precursor protein-dependent mechanism, then penetrates the membrane and is transported to the nucleus and then to autophagosomes. This process is spontaneous or induced by ascorbate depending on the growth-state of the cell. Here, we have explored possible connections between the heparan sulfate traffic route and spermine uptake and delivery in wild-type and Tg2576 mouse fibroblasts. Cells were examined by deconvolution immunofluorescence microscopy. The antibodies used were specific for spermine, glypican-1-derived heparan sulfate, Rab7, nucleolin and a marker for autophagosomes. Endogenous immunostainable spermine was primarily associated with autophagosomes. When spermine synthesis was inhibited, imported spermine appeared in Rab7-positive endosomes. When ascorbate was added, heparan sulfate and spermine were transported to the nucleus where they colocalized with nucleolin. Spermine also appeared in autophagosomes. In a pulse-chase experiment, heparan sulfate and spermine were first arrested in late endosomes by actinomycin D treatment. During the chase, when arrest was abolished, heparan sulfate and spermine were both transported to the nucleus and targeted nucleolin. In amyloid precursor protein<sup>-/-</sup>-fibroblasts, ascorbate failed to induce release of heparan sulfate and spermine remained in the endosomes. We propose that cell surface glypican-1 carries spermine to the endosomes and that the released heparan sulfate carries spermine across the membrane into the cytosol and then to the nucleus.</p>}},
author = {{Cheng, Fang and Fransson, Lars Åke and Mani, Katrin}},
issn = {{0014-4827}},
keywords = {{Amyloid precursor protein; Ascorbate; Autophagosomes; Glypican-1; Heparan sulfate; Nitric oxide; Nucleolin; Spermine}},
language = {{eng}},
month = {{02}},
number = {{2}},
pages = {{133--142}},
publisher = {{Academic Press}},
series = {{Experimental Cell Research}},
title = {{Common traffic routes for imported spermine and endosomal glypican-1-derived heparan sulfate in fibroblasts}},
url = {{http://dx.doi.org/10.1016/j.yexcr.2018.01.029}},
doi = {{10.1016/j.yexcr.2018.01.029}},
volume = {{364}},
year = {{2018}},
}