RPF_MICLU
ID RPF_MICLU Reviewed; 223 AA.
AC O86308;
DT 06-FEB-2013, integrated into UniProtKB/Swiss-Prot.
DT 01-JUN-2002, sequence version 2.
DT 25-MAY-2022, entry version 74.
DE RecName: Full=Resuscitation-promoting factor Rpf;
DE EC=3.-.-.-;
DE Flags: Precursor;
GN Name=rpf;
OS Micrococcus luteus (Micrococcus lysodeikticus).
OC Bacteria; Actinobacteria; Micrococcales; Micrococcaceae; Micrococcus.
OX NCBI_TaxID=1270;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 42-60 AND 81-92,
RP FUNCTION, SUBCELLULAR LOCATION, AND POSSIBLE C-TERMINAL CLEAVAGE.
RC STRAIN=NCIMB 13267 / IAM 14879 / Fleming strain 2665;
RX PubMed=9671779; DOI=10.1073/pnas.95.15.8916;
RA Mukamolova G.V., Kaprelyants A.S., Young D.I., Young M., Kell D.B.;
RT "A bacterial cytokine.";
RL Proc. Natl. Acad. Sci. U.S.A. 95:8916-8921(1998).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], SUBCELLULAR LOCATION, INDUCTION, AND
RP DISRUPTION PHENOTYPE.
RC STRAIN=NCIMB 13267 / IAM 14879 / Fleming strain 2665;
RX PubMed=12410820; DOI=10.1046/j.1365-2958.2002.03183.x;
RA Mukamolova G.V., Turapov O.A., Kazarian K., Telkov M., Kaprelyants A.S.,
RA Kell D.B., Young M.;
RT "The rpf gene of Micrococcus luteus encodes an essential secreted growth
RT factor.";
RL Mol. Microbiol. 46:611-621(2002).
RN [3]
RP FUNCTION AS A MURALYTIC ENZYME, OVEREXPRESSION IN E.COLI, AND MUTAGENESIS
RP OF ASP-48; CYS-53; GLU-54; GLN-72 AND CYS-114.
RC STRAIN=NCIMB 13267 / IAM 14879 / Fleming strain 2665;
RX PubMed=16359320; DOI=10.1111/j.1365-2958.2005.04930.x;
RA Mukamolova G.V., Murzin A.G., Salina E.G., Demina G.R., Kell D.B.,
RA Kaprelyants A.S., Young M.;
RT "Muralytic activity of Micrococcus luteus Rpf and its relationship to
RT physiological activity in promoting bacterial growth and resuscitation.";
RL Mol. Microbiol. 59:84-98(2006).
RN [4]
RP FUNCTION AS A MURALYTIC ENZYME, AND ACTIVITY REGULATION.
RC STRAIN=NCIMB 13267 / IAM 14879 / Fleming strain 2665;
RX PubMed=20016836; DOI=10.1371/journal.pone.0008174;
RA Demina G.R., Makarov V.A., Nikitushkin V.D., Ryabova O.B.,
RA Vostroknutova G.N., Salina E.G., Shleeva M.O., Goncharenko A.V.,
RA Kaprelyants A.S.;
RT "Finding of the low molecular weight inhibitors of resuscitation promoting
RT factor enzymatic and resuscitation activity.";
RL PLoS ONE 4:E8174-E8174(2009).
RN [5]
RP FUNCTION AS A MURALYTIC ENZYME.
RC STRAIN=NCIMB 13267 / IAM 14879 / Fleming strain 2665;
RX PubMed=22864992; DOI=10.1007/s10482-012-9784-1;
RA Nikitushkin V.D., Demina G.R., Shleeva M.O., Kaprelyants A.S.;
RT "Peptidoglycan fragments stimulate resuscitation of 'non-culturable'
RT mycobacteria.";
RL Antonie Van Leeuwenhoek 103:37-46(2013).
CC -!- FUNCTION: Factor that stimulates resuscitation of dormant cells. Has
CC peptidoglycan (PG) hydrolytic activity. Has little to no effect on
CC actively-growing cells. PG fragments could either directly activate the
CC resuscitation pathway of dormant bacteria or serve as a substrate for
CC endogenous Rpf, resulting in low molecular weight products with
CC resuscitation activity. In pM quantities promotes the resuscitation and
CC growth of dormant, nongrowing cells from M.luteus in addition to
CC Mycobacterium tuberculosis, M.avium, M.bovis, M.kansaii and
CC M.smegmatis. Hydrolyzes endogeneous cell walls, peptidoglycan
CC preparations from Mycobacterium tuberculosis and M.smegmatis as well as
CC an artificial lysozyme substrate 4-methylumbelliferyl-beta-D-N,N',N''-
CC triacetylchitotrioside (MUF tri-NAG). Overexpression in E.coli (when
CC the enzyme is targeted to the periplasm) causes cell lysis.
CC {ECO:0000269|PubMed:16359320, ECO:0000269|PubMed:20016836,
CC ECO:0000269|PubMed:22864992, ECO:0000269|PubMed:9671779}.
CC -!- ACTIVITY REGULATION: Activity on the artificial substrate MUF tri-NAG
CC is inhibited by 2-nitrophenylthiocyanates (NPT) compounds.
CC {ECO:0000269|PubMed:20016836}.
CC -!- SUBCELLULAR LOCATION: Secreted. Cell surface. Note=Found in culture
CC supernatant, also accumulates on the cell surface.
CC -!- INDUCTION: RNA is abundant in early lag phase, decreases by early mid-
CC log phase and is undetectable by stationary phase (48 hours growth).
CC Protein expression is slower; detectable by 5 hours, accumulating
CC during early mid-exponential phase and then gradually disappearing by
CC 50 hours growth (late stationary phase) (at protein level).
CC {ECO:0000269|PubMed:12410820}.
CC -!- DOMAIN: The LysM domain is not required for resuscitation activity in
CC vitro; in its absence the protein is active in the fM range.
CC -!- PTM: May be subject to further C-terminal cleavage as the protein
CC identified in gels is smaller than is expected.
CC -!- DISRUPTION PHENOTYPE: Essential, it cannot be disrupted.
CC {ECO:0000269|PubMed:12410820}.
CC -!- SIMILARITY: Belongs to the transglycosylase family. Rpf subfamily.
CC {ECO:0000305}.
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DR EMBL; Z96935; CAB09664.2; -; Genomic_DNA.
DR RefSeq; WP_012750944.1; NZ_NAQZ01000032.1.
DR AlphaFoldDB; O86308; -.
DR SMR; O86308; -.
DR PATRIC; fig|1270.31.peg.1423; -.
DR OMA; VKGGWEK; -.
DR GO; GO:0009986; C:cell surface; IEA:UniProtKB-SubCell.
DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell.
DR GO; GO:0016787; F:hydrolase activity; IEA:UniProtKB-KW.
DR CDD; cd00118; LysM; 1.
DR CDD; cd13925; RPF; 1.
DR Gene3D; 3.10.350.10; -; 1.
DR InterPro; IPR018392; LysM_dom.
DR InterPro; IPR036779; LysM_dom_sf.
DR InterPro; IPR023346; Lysozyme-like_dom_sf.
DR InterPro; IPR010618; RPF.
DR Pfam; PF01476; LysM; 1.
DR Pfam; PF06737; Transglycosylas; 1.
DR SMART; SM00257; LysM; 1.
DR SUPFAM; SSF53955; SSF53955; 1.
DR SUPFAM; SSF54106; SSF54106; 1.
DR PROSITE; PS51782; LYSM; 1.
PE 1: Evidence at protein level;
KW Direct protein sequencing; Hydrolase; Secreted; Signal.
FT SIGNAL 1..41
FT /evidence="ECO:0000269|PubMed:9671779"
FT CHAIN 42..223
FT /note="Resuscitation-promoting factor Rpf"
FT /id="PRO_5000147745"
FT DOMAIN 172..220
FT /note="LysM"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01118"
FT REGION 119..223
FT /note="Not required for resuscitation activity"
FT MUTAGEN 48
FT /note="D->A: <2% MUF tri-NAG hydrolysis, 20% M.luteus cell
FT wall muralytic activity, loss of resuscitation activity;
FT when associated with K-54."
FT /evidence="ECO:0000269|PubMed:16359320"
FT MUTAGEN 53
FT /note="C->K: 6% MUF tri-NAG hydrolysis, 20% M.luteus cell
FT wall muralytic activity, loss of resuscitation activity;
FT when associated with T-114."
FT /evidence="ECO:0000269|PubMed:16359320"
FT MUTAGEN 54
FT /note="E->A: 30% MUF tri-NAG hydrolysis, 25% M.luteus cell
FT wall muralytic activity, significant loss of resuscitation
FT activity."
FT /evidence="ECO:0000269|PubMed:16359320"
FT MUTAGEN 54
FT /note="E->K: <2% MUF tri-NAG hydrolysis, 20% M.luteus cell
FT wall muralytic activity, loss of resuscitation activity;
FT when associated with A-48."
FT /evidence="ECO:0000269|PubMed:16359320"
FT MUTAGEN 54
FT /note="E->Q: 60% MUF tri-NAG hydrolysis, 25% M.luteus cell
FT wall muralytic activity, about 2% resuscitation activity."
FT /evidence="ECO:0000269|PubMed:16359320"
FT MUTAGEN 72
FT /note="Q->K: Little effect on function."
FT /evidence="ECO:0000269|PubMed:16359320"
FT MUTAGEN 114
FT /note="C->T: 6% MUF tri-NAG hydrolysis, 20% M.luteus cell
FT wall muralytic activity, loss of resuscitation activity;
FT when associated with K-53."
FT /evidence="ECO:0000269|PubMed:16359320"
SQ SEQUENCE 223 AA; 23192 MW; 505C564132210705 CRC64;
MDTMTLFTTS ATRSRRATAS IVAGMTLAGA AAVGFSAPAQ AATVDTWDRL AECESNGTWD
INTGNGFYGG VQFTLSSWQA VGGEGYPHQA SKAEQIKRAE ILQDLQGWGA WPLCSQKLGL
TQADADAGDV DATEAAPVAV ERTATVQRQS AADEAAAEQA AAAEQAVVAE AETIVVKSGD
SLWTLANEYE VEGGWTALYE ANKGAVSDAA VIYVGQELVL PQA
