Localization of the type I restriction-modification enzyme EcoKI in the bacterial cell
Copyright 2000 Academic Press..
To localise the type I restriction-modification (R-M) enzyme EcoKI within the bacterial cell, the Hsd subunits present in subcellular fractions were analysed using immunoblotting techniques. The endonuclease (ENase) as well as the methylase (MTase) were found to be associated with the cytoplasmic membrane. HsdR and HsdM subunits produced individually were soluble, cytoplasmic polypeptides and only became membrane-associated when coproduced with the insoluble HsdS subunit. The release of enzyme from the membrane fraction following benzonase treatment indicated a role for DNA in this interaction. Trypsinization of spheroplasts revealed that the HsdR subunit in the assembled ENase was accessible to protease, while HsdM and HsdS, in both ENase and MTase complexes, were fully protected against digestion. We postulate that the R-M enzyme EcoKI is associated with the cytoplasmic membrane in a manner that allows access of HsdR to the periplasmic space, while the MTase components are localised on the inner side of the plasma membrane.
| Medienart: |
Artikel |
|---|
| Erscheinungsjahr: |
2000 |
|---|---|
| Erschienen: |
2000 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:270 |
|---|---|
| Enthalten in: |
Biochemical and biophysical research communications - 270(2000), 1 vom: 02. Apr., Seite 46-51 |
| Sprache: |
Englisch |
|---|
| Beteiligte Personen: |
Holubová, I [VerfasserIn] |
|---|
| Anmerkungen: |
Date Completed 04.05.2000 Date Revised 06.03.2008 published: Print Citation Status MEDLINE |
|---|
| Förderinstitution / Projekttitel: |
|
|---|
| PPN (Katalog-ID): |
NLM106611917 |
|---|
| LEADER | 01000naa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLM106611917 | ||
| 003 | DE-627 | ||
| 005 | 20231222141231.0 | ||
| 007 | tu | ||
| 008 | 231222s2000 xx ||||| 00| ||eng c | ||
| 028 | 5 | 2 | |a pubmed24n0356.xml |
| 035 | |a (DE-627)NLM106611917 | ||
| 035 | |a (NLM)10733903 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 041 | |a eng | ||
| 100 | 1 | |a Holubová, I |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Localization of the type I restriction-modification enzyme EcoKI in the bacterial cell |
| 264 | 1 | |c 2000 | |
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a ohne Hilfsmittel zu benutzen |b n |2 rdamedia | ||
| 338 | |a Band |b nc |2 rdacarrier | ||
| 500 | |a Date Completed 04.05.2000 | ||
| 500 | |a Date Revised 06.03.2008 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright 2000 Academic Press. | ||
| 520 | |a To localise the type I restriction-modification (R-M) enzyme EcoKI within the bacterial cell, the Hsd subunits present in subcellular fractions were analysed using immunoblotting techniques. The endonuclease (ENase) as well as the methylase (MTase) were found to be associated with the cytoplasmic membrane. HsdR and HsdM subunits produced individually were soluble, cytoplasmic polypeptides and only became membrane-associated when coproduced with the insoluble HsdS subunit. The release of enzyme from the membrane fraction following benzonase treatment indicated a role for DNA in this interaction. Trypsinization of spheroplasts revealed that the HsdR subunit in the assembled ENase was accessible to protease, while HsdM and HsdS, in both ENase and MTase complexes, were fully protected against digestion. We postulate that the R-M enzyme EcoKI is associated with the cytoplasmic membrane in a manner that allows access of HsdR to the periplasmic space, while the MTase components are localised on the inner side of the plasma membrane | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 7 | |a Bacterial Proteins |2 NLM | |
| 650 | 7 | |a DNA Restriction-Modification Enzymes |2 NLM | |
| 650 | 7 | |a Escherichia coli Proteins |2 NLM | |
| 650 | 7 | |a HSDS protein, Bacteria |2 NLM | |
| 650 | 7 | |a Membrane Proteins |2 NLM | |
| 650 | 7 | |a DNA modification methylase EcoKI |2 NLM | |
| 650 | 7 | |a EC 2.1.1.- |2 NLM | |
| 650 | 7 | |a HsdM protein, Bacteria |2 NLM | |
| 650 | 7 | |a EC 2.1.1.72 |2 NLM | |
| 650 | 7 | |a Site-Specific DNA-Methyltransferase (Adenine-Specific) |2 NLM | |
| 650 | 7 | |a EC 2.1.1.72 |2 NLM | |
| 650 | 7 | |a Endonucleases |2 NLM | |
| 650 | 7 | |a EC 3.1.- |2 NLM | |
| 650 | 7 | |a DNA Restriction Enzymes |2 NLM | |
| 650 | 7 | |a EC 3.1.21.- |2 NLM | |
| 650 | 7 | |a endodeoxyribonuclease EcoKI |2 NLM | |
| 650 | 7 | |a EC 3.1.21.- |2 NLM | |
| 650 | 7 | |a Deoxyribonucleases, Type I Site-Specific |2 NLM | |
| 650 | 7 | |a EC 3.1.21.3 |2 NLM | |
| 650 | 7 | |a HsdR protein, E coli |2 NLM | |
| 650 | 7 | |a EC 3.1.21.3 |2 NLM | |
| 700 | 1 | |a Vejsadová, S |e verfasserin |4 aut | |
| 700 | 1 | |a Weiserová, M |e verfasserin |4 aut | |
| 700 | 1 | |a Firman, K |e verfasserin |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t Biochemical and biophysical research communications |d 1960 |g 270(2000), 1 vom: 02. Apr., Seite 46-51 |w (DE-627)NLM000000035 |x 1090-2104 |7 nnns |
| 773 | 1 | 8 | |g volume:270 |g year:2000 |g number:1 |g day:02 |g month:04 |g pages:46-51 |
| 912 | |a GBV_USEFLAG_A | ||
| 912 | |a GBV_NLM | ||
| 951 | |a AR | ||
| 952 | |d 270 |j 2000 |e 1 |b 02 |c 04 |h 46-51 | ||