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Characterization of

Deadenylases in Malaria
Transmission
Kevin Hart
Lindner Lab
5/21/15

Outline for Today


Why we care about CCR4
Background
CCR4 Deadenylases
Caf-1 Previous work

Summary: Deadenylases play an important role in the proper


development of Malaria parasites

Plasmodium has a Complex Life Cycle

Original Image By: Maria Mota

PyPuf2-KO results in a loss of spz infectivity


with prolonged salivary gland residence
Down: 129

Up: 513

Lindner et al. Cellular Microbiology. 2013

The CCR4-Not Proteins have different functions


NOT 4
500
0

200
0

FPKM

CCR4-1
10

7.22

CCR4

0.56

0
WT

99

WT

Puf2 KO

Deadenylation
CAF 1
200
0

Not1

50

159

151

WT

Puf2 KO

CAF1 0

24

24

WT

Puf2 KO

50
0

Not2
Not7

0.65

25.74

WT

Puf2 KO

Not5
100

NOT 5

57

4
0

CAF16
2

Puf2 KO

Not4

CAF16
100

WT

N OT 2

CAF40

Puf2 KO

221

Ubiquitination
(-/-) H3K4me3 reduction

CAF40
110

159

WT

Puf2 KO

52

0
WT

Puf2 KO

CCR4-2
4
3
2
1
0

3.37
0.02
WT

Puf2 KO

Modified from Collart, M., & Panasenko, O. (2012). The Ccr4Not complex. Gene, 492(1), 42-53.

Caf1 is the only Not complex protein in


Plasmodium to be characterized
Forward genetic
screen
Piggybac transposon

Caf1 disruption in Pf results in a decreased


parasitemia due to a defect in egress

Conclusions:
KO results in a decrease in proliferation
efficiency
KO results in a severe disruption of merozoite
egress
Premature merozoites bursting RBC
membrane
Attempts to KO in P. berghei unsuccessful

Balu et al. (John Adams) Eukaryotic Cell 2011

Acknowledgements
Lindner Lab
Scott Lindner
Laura Bowman
Steve Griffin
Brooke Kanaskie
Mark Kennedy
Allen Minns
Elyse Munoz
Amanda Reese
Erin Vrana
Mike Walker

Funding Sources
NIAID: 1K22AI101039-01 (SEL)
Penn State Start Up Finds (SEL)