Summary

RMgm-4313
Malaria parasiteP. yoelii
Genotype
DisruptedGene model (rodent): PY17X_1366000; Gene model (P.falciparum): PF3D7_1347500; Gene product: DNA/RNA-binding protein Alba 4 (ALBA4)
Phenotype Gametocyte/Gamete; Oocyst; Sporozoite;
Last modified: 29 August 2017, 10:09
  *RMgm-4313
Successful modificationThe parasite was generated by the genetic modification
The mutant contains the following genetic modification(s) Gene disruption
Reference (PubMed-PMID number) Reference 1 (PMID number) : 28787542
MR4 number
Parent parasite used to introduce the genetic modification
Rodent Malaria ParasiteP. yoelii
Parent strain/lineP. y. yoelii 17XNL
Name parent line/clone Not applicable
Other information parent line
The mutant parasite was generated by
Name PI/ResearcherMuñoz EE, Lindner SE
Name Group/DepartmentDepartment of Biochemistry and Molecular Biology, Center for Malaria Research
Name InstitutePennsylvania State University
CityUniversity Park, PA
CountryUSA
Name of the mutant parasite
RMgm numberRMgm-4313
Principal namepyalba4
Alternative name
Standardized name
Is the mutant parasite cloned after genetic modificationYes
Phenotype
Asexual blood stageNot different from wild type
Gametocyte/GameteNo differences in total gametocytemia, sex ratio, or mature-immature gametocyte ratio; two-fold increase in the number of activated male gametes.
Fertilization and ookineteNot tested
OocystNo significant differences in the prevalence of mosquito infection, oocyst numbers, nor in the total number of sporozoites per infected mosquito (the sum of midgut and salivary gland sporozoites)
A significant effect was observed upon the semi-synchronous egress of sporozoites from oocysts in the midgut and their arrival in the salivary gland; pyalba4- parasites continue to egress from the oocysts over time and are capable of invading the salivary gland and are highly infectious to mice.
SporozoiteA significant effect was observed upon the semi-synchronous egress of sporozoites from oocysts in the midgut and their arrival in the salivary gland; pyalba4- parasites continue to egress from the oocysts over time and are capable of invading the salivary gland and are highly infectious to mice.
Liver stageNot different from wild type
Additional remarks phenotype

Mutant/mutation
The mutant lacks expression of ALBA4 and expresses GFPmut2. The gfpmut2 expression cassette is introduced into the alba4 locus; no information is provided on the promoter driving GFPmut2.

Protein (function)
ALBA proteins are evolutionarily conserved from Archaea and consistently have been found to bind to nucleic acids. Structurally, the ALBA domain resembles the IF3 C-terminal domain, which binds to the small subunit of the prokaryotic ribosome. Together, this strategy provides a straightforward mechanism for ALBA proteins to tether nucleic acids to the ribosome. Plasmodium spp. contain at least four ALBA-domain containing proteins, and have been shown to bind both DNA and RNA. Of these ALBA proteins, ALBA4 is specific to the Apicomplexan lineage and its Chromerid ancestor (40-50%; identity / 60-65% similarity).

Phenotype
No differences in total gametocytemia, sex ratio, or mature-immature gametocyte ratio; two-fold increase in the number of activated male gametes. No significant differences in the prevalence of mosquito infection, oocyst numbers, nor in the total number of sporozoites per infected mosquito (the sum of midgut and salivary gland sporozoites)
A significant effect was observed upon the semi-synchronous egress of sporozoites from oocysts in the midgut and their arrival in the salivary gland; pyalba4- parasites continue to egress from the oocysts over time and are capable of invading the salivary gland and are highly infectious to mice.

Additional information
PyALBA4 localizes to cytosolic puncta throughout the Plasmodium life cycle (see mutant RMgm-4314 expressing GFP-tagged ALBA4).
Evidence is presented that in the absence of ALBA4 i) reduction of specific transcripts in gametocytes ii) increase or reduction of specific transcripts in sporozoites iii) decrease in transcripts in schizonts. The authors conclude based on these observations that 'PyALBA4 plays opposing and large scale roles in regulation distinct transcripts that are relevant to specific stages of development'.
In addition immunoprecipitation studies were performed using GFP-tagged ALBA4 (see mutant RMgm-4314) identifying a number of proteins as putative interacting partners of ALBA4, for example Musashi, Bruno, GBP2, SR1, GAPDH, Aldolase, PABP, a thioredoxin peroxidase, EF1alpha, ribosomal proteins and ALBA1, 2, 3, and 4. The authors conclude: 'the interactions with  ribosome-associated factors are in agreement with previous work that showed that the ALBA domain resembles the IF3-C domain that interacts with the small subunit of the ribosome, and other work that showed that a a member of this complex in yeast (ScDHH1) associates directly with the ribosome.

Other mutants
mutant RMgm-4314 expressing GFP-tagged ALBA4


  Disrupted: Mutant parasite with a disrupted gene
Details of the target gene
Gene Model of Rodent Parasite PY17X_1366000
Gene Model P. falciparum ortholog PF3D7_1347500
Gene productDNA/RNA-binding protein Alba 4
Gene product: Alternative nameALBA4
Details of the genetic modification
Inducable system usedNo
Additional remarks inducable system
Type of plasmid/construct used(Linear) plasmid double cross-over
PlasmoGEM (Sanger) construct/vector usedNo
Modified PlasmoGEM construct/vector usedNo
Plasmid/construct map
Plasmid/construct sequence
Restriction sites to linearize plasmid
Partial or complete disruption of the geneComplete
Additional remarks partial/complete disruption
Selectable marker used to select the mutant parasitehdhfr
Promoter of the selectable markereef1a
Selection (positive) procedurepyrimethamine
Selection (negative) procedureNo
Additional remarks genetic modification
Additional remarks selection procedure
Primer information: Primers used for amplification of the target sequences  Click to view information
Primer information: Primers used for amplification of the target sequences  Click to hide information
Sequence Primer 1
Additional information primer 1
Sequence Primer 2
Additional information primer 2
Sequence Primer 3
Additional information primer 3
Sequence Primer 4
Additional information primer 4
Sequence Primer 5
Additional information primer 5
Sequence Primer 6
Additional information primer 6