Home LiteratureArticle Details
PMID: 9334340 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The phosphatidylinositol transfer protein domain of Drosophila retinal degeneration B protein is essential for photoreceptor cell survival and recovery from light stimulation.

The Journal of cell biology ·Vol. 139 ·No. 2 ·1997-10-20 ·Pages 351-63

Milligan SC, Alb JG, Elagina RB, Bankaitis VA, Hyde DR

Abstract

The Drosophila retinal degeneration B (rdgB) gene encodes an integral membrane protein involved in phototransduction and prevention of retinal degeneration. RdgB represents a nonclassical phosphatidylinositol transfer protein (PITP) as all other known PITPs are soluble polypeptides. Our data demonstrate roles for RdgB in proper termination of the phototransduction light response and dark recovery of the photoreceptor cells. Expression of RdgB's PITP domain as a soluble protein (RdgB-PITP) in rdgB2 mutant flies is sufficient to completely restore the wild-type electrophysiological light response and prevent the degeneration. However, introduction of the T59E mutation, which does not affect RdgB-PITP's phosphatidylinositol (PI) and phosphatidycholine (PC) transfer in vitro, into the soluble (RdgB-PITP-T59E) or full-length (RdgB-T59E) proteins eliminated rescue of retinal degeneration in rdgB2 flies, while the light response was partially maintained. Substitution of the rat brain PITPalpha, a classical PI transfer protein, for RdgB's PITP domain (PITPalpha or PITPalpha-RdgB chimeric protein) neither restored the light response nor maintained retinal integrity when expressed in rdgB2 flies. Therefore, the complete repertoire of essential RdgB functions resides in RdgB's PITP domain, but other PITPs possessing PI and/or PC transfer activity in vitro cannot supplant RdgB function in vivo. Expression of either RdgB-T59E or PITPalpha-RdgB in rdgB+ flies produced a dominant retinal degeneration phenotype. Whereas RdgB-T59E functioned in a dominant manner to significantly reduce steady-state levels of rhodopsin, PITPalpha-RdgB was defective in the ability to recover from prolonged light stimulation and caused photoreceptor degeneration through an unknown mechanism. This in vivo analysis of PITP function in a metazoan system provides further insights into the links between PITP dysfunction and an inherited disease in a higher eukaryote.

MeSH Terms
Aging Animals Brain/metabolism Carrier Proteins/chemistry,metabolism Cloning, Molecular Drosophila/genetics,physiology Drosophila Proteins Electrophysiology/methods Eye Proteins Light Membrane Proteins/chemistry,metabolism Phosphatidylinositols Phospholipid Transfer Proteins Photic Stimulation Photoreceptor Cells, Invertebrate/cytology,pathology,physiology Point Mutation Polymerase Chain Reaction Rats Recombinant Proteins/chemistry,metabolism Threonine
Chemicals
Carrier Proteins Drosophila Proteins Eye Proteins Membrane Proteins Phosphatidylinositols Phospholipid Transfer Proteins Recombinant Proteins rdgB protein, Drosophila Threonine
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Milligan S C
Department of Biological Sciences, University of Notre Dame, Notre Dame, Indiana 46556, USA.
Alb J G
Elagina R B
Bankaitis V A
Hyde D R
References (46)
46 references, click to expand
  1. A phosphatidylinositol transfer protein controls the phosphatidylcholine content of yeast Golgi membranes.
    J Cell Biol. 1994 Feb;124(3):273-87 PMID: 8294512
  2. A Drosophila mutant defective in extracellular calcium-dependent photoreceptor deactivation and rapid desensitization.
    Nature. 1991 Nov 21;354(6350):230-2 PMID: 1961249
  3. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  4. Mammalian homolog of Drosophila retinal degeneration B rescues the mutant fly phenotype.
    J Neurosci. 1997 Aug 1;17(15):5881-90 PMID: 9221785
  5. Chemically induced retinal degeneration in the rdgB (retinal degeneration B) mutant of Drosophila.
    Vis Neurosci. 1989;2(6):541-51 PMID: 2518632
  6. Phospholipid transfer activity is relevant to but not sufficient for the essential function of the yeast SEC14 gene product.
    EMBO J. 1993 Dec;12(12):4775-84 PMID: 8223486
  7. Hereditary retinal degeneration in Drosophila melanogaster. A mutant defect associated with the phototransduction process.
    J Gen Physiol. 1977 Mar;69(3):261-91 PMID: 139462
  8. Signal transduction. Phospholipids in action.
    Nature. 1995 Jan 19;373(6511):194-5 PMID: 7816126
  9. Immunolocalization of a Drosophila phosphatidylinositol transfer protein (rdgB) in normal and rdgA mutant photoreceptor cells with special reference to the subrhabdomeric cisternae.
    J Electron Microsc (Tokyo). 1994 Aug;43(4):183-9 PMID: 7996077
  10. The Saccharomyces cerevisiae SEC14 gene encodes a cytosolic factor that is required for transport of secretory proteins from the yeast Golgi complex.
    J Cell Biol. 1989 Apr;108(4):1271-81 PMID: 2466847
  11. Essential role for diacylglycerol in protein transport from the yeast Golgi complex.
    Nature. 1997 May 1;387(6628):101-5 PMID: 9139830
  12. Rhodopsin plays an essential structural role in Drosophila photoreceptor development.
    Development. 1995 Dec;121(12):4359-70 PMID: 8575336
  13. An essential role for phosphatidylinositol transfer protein in phospholipase C-mediated inositol lipid signaling.
    Cell. 1993 Sep 10;74(5):919-28 PMID: 8374957
  14. Isolation and characterization of the Drosophila retinal degeneration B (rdgB) gene.
    Genetics. 1991 Apr;127(4):761-8 PMID: 1903119
  15. Rapid and efficient site-specific mutagenesis without phenotypic selection.
    Methods Enzymol. 1987;154:367-82 PMID: 3323813
  16. Phospholipid transfer proteins: a biological debut.
    Trends Cell Biol. 1991 Jul;1(1):30-4 PMID: 14731807
  17. Mutant rat phosphatidylinositol/phosphatidylcholine transfer proteins specifically defective in phosphatidylinositol transfer: implications for the regulation of phospholipid transfer activity.
    Proc Natl Acad Sci U S A. 1995 Sep 12;92(19):8826-30 PMID: 7568025
  18. rdgE: a novel retinal degeneration mutation in Drosophila melanogaster.
    Genetics. 1996 Sep;144(1):127-38 PMID: 8878679
  19. Maturation of major Drosophila rhodopsin, ninaE, requires chromophore 3-hydroxyretinal.
    Neuron. 1993 Jun;10(6):1113-9 PMID: 8318232
  20. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  21. An essential role for a phospholipid transfer protein in yeast Golgi function.
    Nature. 1990 Oct 11;347(6293):561-2 PMID: 2215682
  22. The Saccharomyces cerevisiae phosphatidylinositol-transfer protein effects a ligand-dependent inhibition of choline-phosphate cytidylyltransferase activity.
    Proc Natl Acad Sci U S A. 1995 Jan 3;92(1):112-6 PMID: 7816798
  23. Immunoelectron microscopic study of the opsin distribution in the photoreceptor cells of Drosophila melanogaster.
    J Electron Microsc (Tokyo). 1991 Jun;40(3):187-92 PMID: 1838762
  24. The Drosophila ninaE gene encodes an opsin.
    Cell. 1985 Apr;40(4):839-50 PMID: 2985266
  25. Phospholipid transfer proteins.
    Annu Rev Biochem. 1991;60:73-99 PMID: 1883207
  26. Photoreceptor deactivation and retinal degeneration mediated by a photoreceptor-specific protein kinase C.
    Science. 1991 Dec 6;254(5037):1478-84 PMID: 1962207
  27. Structure of the subrhabdomeric cisternae in the photoreceptor cells of Drosophila melanogaster.
    J Neurocytol. 1989 Feb;18(1):87-93 PMID: 2496206
  28. Drosophila mutants with reduced rhodopsin content.
    Symp Soc Exp Biol. 1983;36:477-501 PMID: 6399788
  29. Isolation of a putative phospholipase C gene of Drosophila, norpA, and its role in phototransduction.
    Cell. 1988 Aug 26;54(5):723-33 PMID: 2457447
  30. Isolation and structure of a rhodopsin gene from D. melanogaster.
    Cell. 1985 Apr;40(4):851-8 PMID: 2580638
  31. Degeneration of photoreceptors in rhodopsin mutants of Drosophila.
    J Neurobiol. 1992 Aug;23(6):605-26 PMID: 1431838
  32. Phospholipid metabolism and membrane dynamics.
    Curr Opin Cell Biol. 1996 Aug;8(4):534-41 PMID: 8791444
  33. Phosphatidylinositol transfer protein required for ATP-dependent priming of Ca(2+)-activated secretion.
    Nature. 1993 Dec 9;366(6455):572-5 PMID: 8255295
  34. Calcium-sequestering smooth endoplasmic reticulum in retinula cells of the blowfly.
    J Ultrastruct Res. 1982 Nov;81(2):240-8 PMID: 7143549
  35. The biology of vision of Drosophila.
    Proc Natl Acad Sci U S A. 1996 Jan 23;93(2):571-6 PMID: 8570597
  36. The Drosophila dgq gene encodes a G alpha protein that mediates phototransduction.
    Neuron. 1994 Nov;13(5):1143-57 PMID: 7946351
  37. Localization of Drosophila retinal degeneration B, a membrane-associated phosphatidylinositol transfer protein.
    J Cell Biol. 1993 Sep;122(5):1013-22 PMID: 8354691
  38. The vibrator mutation causes neurodegeneration via reduced expression of PITP alpha: positional complementation cloning and extragenic suppression.
    Neuron. 1997 May;18(5):711-22 PMID: 9182797
  39. The gene encoding the phosphatidylinositol transfer protein is essential for cell growth.
    J Biol Chem. 1990 Mar 15;265(8):4711-7 PMID: 2407740
  40. A role for phosphatidylinositol transfer protein in secretory vesicle formation.
    Nature. 1995 Oct 12;377(6549):544-7 PMID: 7566155
  41. Phorbol ester induces photoreceptor-specific degeneration in a Drosophila mutant.
    Proc Natl Acad Sci U S A. 1990 Jan;87(1):113-7 PMID: 2296574
  42. Analysis of the promoter of the ninaE opsin gene in Drosophila melanogaster.
    Genetics. 1987 Aug;116(4):565-78 PMID: 3040524
  43. Protein kinase C is required for light adaptation in Drosophila photoreceptors.
    Nature. 1993 Jun 17;363(6430):634-7 PMID: 8510756
  44. Phosphatidylinositol transfer protein dictates the rate of inositol trisphosphate production by promoting the synthesis of PIP2.
    Curr Biol. 1995 Jul 1;5(7):775-83 PMID: 7583124
  45. ATP-dependent inositide phosphorylation required for Ca(2+)-activated secretion.
    Nature. 1995 Mar 9;374(6518):173-7 PMID: 7877690
  46. Regulation of PLC-mediated signalling in vivo by CDP-diacylglycerol synthase.
    Nature. 1995 Jan 19;373(6511):216-22 PMID: 7816135
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.139.2.351
Published
1997-10-20
Pages
351-63
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2139788
Subset
IM
Grants
NEI NIH HHS · EY08058 · United States
NIGMS NIH HHS · GM44530 · United States
External Links
PubMed source DOI: 10.1083/jcb.139.2.351 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2