---
OA_place: publisher
OA_type: hybrid
PlanS_conform: '1'
_id: '20404'
abstract:
- lang: eng
  text: Collagens are fundamental components of extracellular matrices, requiring
    precise intracellular post-translational modifications for proper function. Among
    the modifications, prolyl 4-hydroxylation is critical to stabilise the collagen
    triple helix. In humans, this reaction is mediated by collagen prolyl 4-hydroxylases
    (P4Hs). While humans possess three genes encoding these enzymes (P4H⍺s), Drosophila
    melanogaster harbour at least 26 candidates for collagen P4H⍺s despite its simple
    genome, and it is poorly understood which of them are actually working on collagen
    in the fly. In this study, we addressed this question by carrying out thorough
    bioinformatic and biochemical analyses. We demonstrate that among the 26 potential
    collagen P4H⍺s, PH4⍺EFB shares the highest homology with vertebrate collagen P4H⍺s.
    Furthermore, while collagen P4Hs and their substrates must exist in the same cells,
    our transcriptomic analyses at the tissue and single cell levels showed a global
    co-expression of PH4⍺EFB but not the other P4H⍺-related genes with the collagen
    IV genes. Moreover, expression of PH4⍺EFB during embryogenesis was found to precede
    that of collagen IV, presumably enabling efficient collagen modification by PH4⍺EFB.
    Finally, biochemical assays confirm that PH4⍺EFB binds collagen, supporting its
    direct role in collagen IV modification. Collectively, we identify PH4⍺EFB as
    the primary and potentially constitutive prolyl 4-hydroxylase responsible for
    collagen IV biosynthesis in Drosophila. Our findings highlight the remarkably
    simple nature of Drosophila collagen IV biosynthesis, which may serve as a blueprint
    for defining the minimal requirements for collagen engineering.
acknowledgement: "This project was supported by the All May See Foundation 7031,182
  to YI, the Louisiana Board of Regents Support Fund: Research Competitiveness Subprogram
  to MAT, Austrian science fund (FWF) as part of the SFB Meiosis consortium FWF SFB
  F88-10 to Beatriz Vicoso (supported ME), American Heart Association 16POST2726018
  and American Cancer Society 132,123-PF-18–025–01-CSM postdoctoral fellowships to
  ALZ, National Institutes of Health R01 GM136961 and R35 GM148485 to SH-B, and the
  Academy of Medical Sciences/the Wellcome Trust/ the Government Department of Business,
  Energy and Industrial Strategy/the British Heart Foundation/Diabetes UK Springboard
  Award SBF008\\1115 to YM. \r\nComputational analyses of single-nucleus transcriptome
  data were performed on the high performance computer (HPC) at Bournemouth University,
  the HPC at Institute of Science and Technology Austria, and the high-performance
  computational resources provided by the Louisiana Optical Network Infrastructure
  (http://www.loni.org). The authors are grateful to the researchers who published
  the transcriptome datasets [48,49,52,55] that became the essential bases for this
  study, to FlyBase for curating the datasets in an easily accessible format, and
  the Drosophila Genomics Resource Center (DGRC), supported by NIH grant 2P40OD010949,
  for providing the D17 cell line used in this research. The authors thank Kristian
  Koski (University of Oulu, Finland) for crucial advice on the domain structure of
  collagen P4H⍺s, and Ryusuke Niwa and Ryo Hoshino (University of Tsukuba, Japan)
  for helpful discussions on SP."
article_processing_charge: Yes (in subscription journal)
article_type: original
author:
- first_name: Yoshihiro
  full_name: Ishikawa, Yoshihiro
  last_name: Ishikawa
- first_name: Melissa A
  full_name: Toups, Melissa A
  id: 4E099E4E-F248-11E8-B48F-1D18A9856A87
  last_name: Toups
  orcid: 0000-0002-9752-7380
- first_name: Marwan N
  full_name: Elkrewi, Marwan N
  id: 0B46FACA-A8E1-11E9-9BD3-79D1E5697425
  last_name: Elkrewi
  orcid: 0000-0002-5328-7231
- first_name: Allison L.
  full_name: Zajac, Allison L.
  last_name: Zajac
- first_name: Sally
  full_name: Horne-Badovinac, Sally
  last_name: Horne-Badovinac
- first_name: Yutaka
  full_name: Matsubayashi, Yutaka
  last_name: Matsubayashi
citation:
  ama: Ishikawa Y, Toups MA, Elkrewi MN, Zajac AL, Horne-Badovinac S, Matsubayashi
    Y. Evidence for the major role of PH4⍺EFB in the prolyl 4-hydroxylation of Drosophila
    collagen IV. <i>Matrix Biology</i>. 2025;141(11):101-113. doi:<a href="https://doi.org/10.1016/j.matbio.2025.09.002">10.1016/j.matbio.2025.09.002</a>
  apa: Ishikawa, Y., Toups, M. A., Elkrewi, M. N., Zajac, A. L., Horne-Badovinac,
    S., &#38; Matsubayashi, Y. (2025). Evidence for the major role of PH4⍺EFB in the
    prolyl 4-hydroxylation of Drosophila collagen IV. <i>Matrix Biology</i>. Springer
    Nature. <a href="https://doi.org/10.1016/j.matbio.2025.09.002">https://doi.org/10.1016/j.matbio.2025.09.002</a>
  chicago: Ishikawa, Yoshihiro, Melissa A Toups, Marwan N Elkrewi, Allison L. Zajac,
    Sally Horne-Badovinac, and Yutaka Matsubayashi. “Evidence for the Major Role of
    PH4⍺EFB in the Prolyl 4-Hydroxylation of Drosophila Collagen IV.” <i>Matrix Biology</i>.
    Springer Nature, 2025. <a href="https://doi.org/10.1016/j.matbio.2025.09.002">https://doi.org/10.1016/j.matbio.2025.09.002</a>.
  ieee: Y. Ishikawa, M. A. Toups, M. N. Elkrewi, A. L. Zajac, S. Horne-Badovinac,
    and Y. Matsubayashi, “Evidence for the major role of PH4⍺EFB in the prolyl 4-hydroxylation
    of Drosophila collagen IV,” <i>Matrix Biology</i>, vol. 141, no. 11. Springer
    Nature, pp. 101–113, 2025.
  ista: Ishikawa Y, Toups MA, Elkrewi MN, Zajac AL, Horne-Badovinac S, Matsubayashi
    Y. 2025. Evidence for the major role of PH4⍺EFB in the prolyl 4-hydroxylation
    of Drosophila collagen IV. Matrix Biology. 141(11), 101–113.
  mla: Ishikawa, Yoshihiro, et al. “Evidence for the Major Role of PH4⍺EFB in the
    Prolyl 4-Hydroxylation of Drosophila Collagen IV.” <i>Matrix Biology</i>, vol.
    141, no. 11, Springer Nature, 2025, pp. 101–13, doi:<a href="https://doi.org/10.1016/j.matbio.2025.09.002">10.1016/j.matbio.2025.09.002</a>.
  short: Y. Ishikawa, M.A. Toups, M.N. Elkrewi, A.L. Zajac, S. Horne-Badovinac, Y.
    Matsubayashi, Matrix Biology 141 (2025) 101–113.
date_created: 2025-09-28T22:01:26Z
date_published: 2025-11-01T00:00:00Z
date_updated: 2026-01-05T13:09:08Z
day: '01'
ddc:
- '570'
department:
- _id: BeVi
doi: 10.1016/j.matbio.2025.09.002
external_id:
  isi:
  - '001583892100002'
  pmid:
  - '40946811'
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has_accepted_license: '1'
intvolume: '       141'
isi: 1
issue: '11'
language:
- iso: eng
license: https://creativecommons.org/licenses/by/4.0/
month: '11'
oa: 1
oa_version: Published Version
page: 101-113
pmid: 1
project:
- _id: 34ae1506-11ca-11ed-8bc3-c14f4c474396
  grant_number: F8810
  name: The highjacking of meiosis for asexual reproduction
publication: Matrix Biology
publication_identifier:
  eissn:
  - 1569-1802
  issn:
  - 0945-053X
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
scopus_import: '1'
status: public
title: Evidence for the major role of PH4⍺EFB in the prolyl 4-hydroxylation of Drosophila
  collagen IV
tmp:
  image: /images/cc_by.png
  legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
  name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
  short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 141
year: '2025'
...