EC Accepted Manuscript Posted Online 10 April 2015 Eukaryotic Cell doi:10.1128/EC.00010-15 Copyright © 2015, American Society for Microbiology. All Rights Reserved.

1



2

,PSDFWRISURWHLQSDOPLWR\ODWLRQRQWKHYLUXOHQFHSRWHQWLDORICryptococcus

3

neoformans

4 5 6

&RQQLH%1LFKROV.\OD62VW'D\WRQ3*URJDQ.DLOD3LDQDOWR6KLULQ

7

+DVDQDQG-$QGUHZ$OVSDXJK

8



9



10

'HSDUWPHQWVRI0HGLFLQHDQG0ROHFXODU*HQHWLFV0LFURELRORJ\

11

'XNH8QLYHUVLW\6FKRRORI0HGLFLQH

12

'XUKDP1&

13



14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34

        

&RUUHVSRQGHQFHWR'U$OVSDXJK 'XNH+RVSLWDO6RXWK '80& 'XNH8QLYHUVLW\0HGLFDO&HQWHU 'XUKDP1& 7HOH)D[ (PDLODQGUHZDOVSDXJK#GXNHHGX   UXQQLQJWLWOH3DOPLWR\ODWLRQLQC. neoformans NH\ZRUGVYLUXOHQFHVLJQDOWUDQVGXFWLRQIXQJXVPRUSKRJHQHVLV6 DF\OWUDQVIHUDVH





35

$EVWUDFW

36

7KHORFDOL]DWLRQDQGVSHFLDOL]HGIXQFWLRQRI5DVOLNHSURWHLQVDUHODUJHO\

37

GHWHUPLQHGE\SRVWWUDQVODWLRQDOSURFHVVLQJHYHQWV,QDKLJKO\UHJXODWHG

38

SURFHVVSDOPLWR\OJURXSVPD\EHDGGHGWR&WHUPLQDOF\VWHLQHUHVLGXHV

39

WDUJHWLQJWKHVHSURWHLQVWRVSHFLILFPHPEUDQHV,QWKHKXPDQIXQJDOSDWKRJHQ

40

Cryptococcus neoformans5DVSURWHLQSDOPLWR\ODWLRQLVHVVHQWLDOIRUJURZWKDW

41

KLJKWHPSHUDWXUHEXWLVGLVSHQVDEOHIRUVH[XDOGLIIHUHQWLDWLRQ5DV

42

SDOPLWR\ODWLRQLVDOVRUHTXLUHGIRUORFDOL]DWLRQRIWKLVSURWHLQRQWKHSODVPD

43

PHPEUDQH7RJHWKHUWKHVHUHVXOWVVXSSRUWDPRGHOLQZKLFKVSHFLILF5DV

44

IXQFWLRQVDUHPHGLDWHGIURPGLIIHUHQWVXEFHOOXODUORFDWLRQV:HWKHUHIRUH

45

K\SRWKHVL]HWKDWSURWHLQVWKDWDFWLYDWH5DVRUPHGLDWH5DVORFDOL]DWLRQWRWKH

46

SODVPDPHPEUDQHZLOOEHLPSRUWDQWIRUC. neoformansSDWKRJHQHVLV7RIXUWKHU

47

FKDUDFWHUL]HWKH5DVVLJQDOLQJFDVFDGHPHGLDWLQJKLJKWHPSHUDWXUHJURZWKZH

48

KDYHLGHQWLILHGDIDPLO\RISURWHLQ6DF\OWUDQVIHUDVHV3$7V HQ]\PHVWKDW

49

PHGLDWHSDOPLWR\ODWLRQLQWKHC. neoformansJHQRPHGDWDEDVH'HOHWLRQVWUDLQV

50

IRUHDFKFDQGLGDWHJHQHZHUHJHQHUDWHGE\KRPRJHQRXVUHFRPELQDWLRQDQG

51

HDFKPXWDQWVWUDLQZDVDVVHVVHGIRU5DVPHGLDWHGSKHQRW\SHVLQFOXGLQJKLJK

52

WHPSHUDWXUHJURZWKPRUSKRJHQHVLVDQGVH[XDOGHYHORSPHQW:HIRXQGWKDWIXOO

53

5DVSDOPLWR\ODWLRQDQGIXQFWLRQUHTXLUHGRQHSDUWLFXODU3$73IDDQGGHOHWLRQ

54

RIWKHPFA4JHQHLQ C. neoformansUHVXOWHGLQDOWHUHG5DVORFDOL]DWLRQWR

55

PHPEUDQHVLPSDLUHGJURZWKDWƒ&DQGUHGXFHGYLUXOHQFH

56





57

,QWURGXFWLRQ

58



59

PRLHWLHVWRSURWHLQV7KLVSURFHVVRISRVWWUDQVODWLRQDOPRGLILFDWLRQLVFDWDO\]HG

60

E\WKHSURWHLQ6DF\OWUDQVIHUDVH3$7 HQ]\PHVWKURXJKWKHWKLRHVWHULILFDWLRQRI

61

DF\VWHLQHVLGHFKDLQJURXS%\WKHDGGLWLRQRIWKLVK\GURSKRELFSDOPLWR\OJURXS

62

SURWHLQVRIWHQEHFRPHDVVRFLDWHGZLWKPHPEUDQHV+RZHYHULQFRQWUDVWWR

63

FRQVWLWXWLYHSURWHLQPRGLILFDWLRQVVXFKDVSUHQ\ODWLRQSDOPLWR\ODWLRQLVRIWHQ

64

UHJXODWHGDQGUHYHUVLEOHDOORZLQJIRUG\QDPLFFKDQJHVLQSDWWHUQVRIVXEFHOOXODU

65

SURWHLQORFDOL]DWLRQ  

66

6SDOPLWR\ODWLRQLQYROYHVWKHUHYHUVLEOHDWWDFKPHQWRISDOPLWLFDFLG

3$7JHQHVZHUHILUVWLGHQWLILHGLQWKHEXGGLQJ\HDVWS. cerevisiaeDVWKH

67

HQ]\PHVUHVSRQVLEOHIRUWKHSDOPLWR\ODWLRQRI5DV(UI DQG